Sample records for copper nitrite reductase

The copper-containing nitritereductase from Alcaligenes faecalis S-6 was found to catalyze the oxidation of nitric oxide to nitrite, the reverse of its physiological reaction. Thermodynamic and kinetic constants with the physiological electron donor pseudoazurin were determined for both directions

Many properties of copper-containing nitritereductase are pH-dependent, such as gene expression, enzyme activity, and substrate affinity. Here we use x-ray diffraction to investigate the structural basis for the pH dependence of activity and nitrite affinity by examining the type 2 copper site and

The intramolecular electron transfer (ET) between the type 1 Cu(I) and the type 2 Cu(II) sites of Alcaligenes xylosoxidans dissimilatory nitritereductase (AxNiR) has been studied in order to compare it with the analogous process taking place in ascorbate oxidase (AO). This internal process...

We studied the electrochemical behavior of the redox metalloenzyme coppernitritereductase (CNiR, Achromobacter xylosoxidans) immobilized on a Au(111)‐electrode surface modified by a self‐assembled cysteamine molecular monolayer (SAM) using a combination of cyclic voltammetry and electrochemically......‐controlled atomic force microscopy (in situ AFM). The enzyme showed no voltammetric signals in the absence of nitrite substrate, whereas a strong reductive electrocatalytic signal appeared in the presence of nitrite. Such a pattern is common in protein film and monolayer voltammetry and points to conformational...... in the presence of nitrite. No change in size was observed in the absence of nitrite over the same potential range. The enzyme size variation is suggested to offer clues to the broadly observed substrate triggering in metalloenzyme monolayer voltammetry....

Nitric oxide (NO) as a cellular signaling molecule and vasodilator regulates a range of physiological and pathological processes. Nitrite (NO2 (-)) is recycled in vivo to generate nitric oxide, particularly in physiologic hypoxia and ischemia. The cytochrome c oxidase binuclear heme a 3/CuB active site is one entity known to be responsible for conversion of cellular nitrite to nitric oxide. We recently reported that a partially reduced heme/copper assembly reduces nitrite ion, producing nitric oxide; the heme serves as the reductant and the cupric ion provides a Lewis acid interaction with nitrite, facilitating nitrite (N-O) bond cleavage (Hematian et al., J. Am. Chem. Soc. 134:18912-18915, 2012). To further investigate this nitritereductase chemistry, copper(II)-nitrito complexes with tridentate and tetradentate ligands were used in this study, where either O,O'-bidentate or O-unidentate modes of nitrite binding to the cupric center are present. To study the role of the reducing ability of the ferrous heme center, two different tetraarylporphyrinate-iron(II) complexes, one with electron-donating para-methoxy peripheral substituents and the other with electron-withdrawing 2,6-difluorophenyl substituents, were used. The results show that differing modes of nitrite coordination to the copper(II) ion lead to differing kinetic behavior. Here, also, the ferrous heme is in all cases the source of the reducing equivalent required to convert nitrite to nitric oxide, but the reduction ability of the heme center does not play a key role in the observed overall reaction rate. On the basis of our observations, reaction mechanisms are proposed and discussed in terms of heme/copper heterobinuclear structures.

Nitritereductase (NiR) is an enzyme that uses type 1 and type 2 copper sites to reduce nitrite to nitric oxide during bacterial denitrification. A copper-nitrosyl intermediate is a proposed, yet poorly characterized feature of the NiR catalytic cycle. This intermediate is formally described as Cu(I)-NO{sup +} and is proposed to be formed at the type 2 copper site after nitrite binding and electron transfer from the type 1 copper site. In this study, copper-nitrosyl complexes were formed by prolonged exposure of exogenous NO to crystals of wild-type and two variant forms of NiR from Alcaligenes faecalis (AfNiR), and the structures were determined to 1.8 {angstrom} or better resolution. Exposing oxidized wild-type crystals to NO results in the reverse reaction and formation of nitrite that remains bound at the active site. In a type 1 copper site mutant (H145A) that is incapable of electron transfer to the type 2 site, the reverse reaction is not observed. Instead, in both oxidized and reduced H145A crystals, NO is observed bound in a side-on manner to the type 2 copper. In AfNiR, Asp98 forms hydrogen bonds to both substrate and product bound to the type 2 Cu. In the D98N variant, NO is bound side-on but is more disordered when observed for the wild-type enzyme. The solution EPR spectra of the crystallographically characterized NiR-NO complexes indicate the presence of an oxidized type 2 copper site and thus are interpreted as resulting from stable copper-nitrosyls and formally assigned as Cu(II)-NO{sup -}. A reaction scheme in which a second NO molecule is oxidized to nitrite can account for the formation of a CuD-NO{sup -} species after exposure of the oxidized H145A variant to NO gas.

Using density functional theory, we studied denitrification reaction mechanisms of copper adducts of tris(pyrazolyl)methane and hydrotris(pyrazolyl)borate models of a coppernitritereductase (Cu-NiR), and herein propose several possible reaction pathways, including some parts that have never been examined previously. Because electron and proton transfer reactions participate in the enzymatic cycles of Cu-NiR, the Gibbs energy of a proton in solution, G(H(+)), and the redox potential, Eredox, of the model Cu-NiR are also evaluated. Although the pathway where a nitrite is provided as HNO2 is energetically preferable, a well-known reaction pathway passing through the resting state with an active site occupied by a water molecule where nitrite is provided as NO2(-) is the main recognized pathway under normal conditions. These features do not change whether the electron transfer occurs before production of NO or not. However, our results suggest that the pathway involving HNO2 might become dominant under low pH conditions in conjunction with experimental results.

This thesis is concerned with the synthesis of a series of novel pyridine containing ligands and their copper co-ordination chemistry. The aim was to design ligands which would produce copper complexes which model the active sites within certain copper-containing NitriteReductase enzymes. The first chapter reviews previous work in this area and details the promising nature of pyridine-containing ligands. The remainder of this thesis is concerned with the synthesis and characterisation of some novel pyridine-containing ligands and their copper chemistry. The synthetic routes developed during this work enabled tris(pyrid-2-yl)methylamine ligands to be produced and studied which were tripodal in form but which had a primary amine group at the cap which could be further elaborated. Additional substituents were also placed on the pyridine rings to investigate their impact on the chemistry of their copper complexes. These ligands showed a variety, counter ion dependent chemistry. The structures of number of the co...

We have studied the adsorption and electrocatalysis of the redox metalloenzyme blue coppernitritereductase from Achromobacter xylosoxidans (AxCuNiR) on single-crystal Au(111)-electrode surfaces modified by a self-assembled monolayer of cysteamine. A combination of cyclic voltammetry and in situ...... biotechnology at the monolayer and toward the single-molecule level....

We have studied the adsorption and electrocatalysis of the redox metalloenzyme blue coppernitritereductase from Achromobacter xylosoxidans (AxCuNiR) on single-crystal Au(111)-electrode surfaces modified by a self-assembled monolayer of cysteamine. A combination of cyclic voltammetry and in situ...

Relating individual protein crystal structures to an enzyme mechanism remains a major and challenging goal for structural biology. Serial crystallography using multiple crystals has recently been reported in both synchrotron-radiation and X-ray free-electron laser experiments. In this work, serial crystallography was used to obtain multiple structures serially from one crystal (MSOX) to study in crystallo enzyme catalysis. Rapid, shutterless X-ray detector technology on a synchrotron MX beamline was exploited to perform low-dose serial crystallography on a single coppernitritereductase crystal, which survived long enough for 45 consecutive 100 K X-ray structures to be collected at 1.07-1.62 Å resolution, all sampled from the same crystal volume. This serial crystallography approach revealed the gradual conversion of the substrate bound at the catalytic type 2 Cu centre from nitrite to nitric oxide, following reduction of the type 1 Cu electron-transfer centre by X-ray-generated solvated electrons. Significant, well defined structural rearrangements in the active site are evident in the series as the enzyme moves through its catalytic cycle, namely nitrite reduction, which is a vital step in the global denitrification process. It is proposed that such a serial crystallography approach is widely applicable for studying any redox or electron-driven enzyme reactions from a single protein crystal. It can provide a 'catalytic reaction movie' highlighting the structural changes that occur during enzyme catalysis. The anticipated developments in the automation of data analysis and modelling are likely to allow seamless and near-real-time analysis of such data on-site at some of the powerful synchrotron crystallographic beamlines.

The efficient application of nitrogenous fertilizers is urgently required, as their excessive and inefficient use is causing substantial economic loss and environmental pollution. A significant amount of applied nitrogen in agricultural soils is lost as nitrous oxide (N2O) in the environment due to the microbial denitrification process. The widely distributed fungus Fusarium oxysporum is a major denitrifier in agricultural soils and its denitrification activity could be targeted to reduce nitrogen loss in the form of N2O from agricultural soils. Here, we report the discovery of first small molecule inhibitors of coppernitritereductase (NirK) from F. oxysporum, which is a key enzyme in the fungal denitrification process. The inhibitors were discovered by a hierarchical in silico screening approach consisting of pharmacophore modeling and molecular docking. In vitro evaluation of F. oxysporum NirK activity revealed several pyrimidone and triazinone based compounds with potency in the low micromolar range. Some of these compounds suppressed the fungal denitrification in vivo as well. The compounds reported here could be used as starting points for the development of nitrogenous fertilizer supplements and coatings as a means to prevent nitrogen loss by targeting fungal denitrification.

Enzymatic reactions involving redox processes are highly sensitive to the local electrostatic environment. Despite considerable effort, the complex interactions among different influential factors in native proteins impede progress toward complete understanding of the structure-function relationship. Of particular interest is the type 2 copper center Cu(His)3, which may act as an electron transfer center in peptidylglycine α-hydroxylating monooxygenase (PHM) or a catalytic center in coppernitritereductase (CuNiR). A de novo design strategy is used to probe the effect of modifying charged amino acid residues around, but not directly bound to, a Cu(His)3 center embedded in three-stranded coiled coils (TRI-H)3 [TRI-H = Ac-G WKALEEK LKALEEK LKALEEK HKALEEK G-NH2]. Specifically, the peptide TRI-EH (=TRI-HK22E) alters an important lysine to glutamate just above the copper binding center. With a series of TRI-EH peptides mutated below the metal center, we use a variety of spectroscopies (EPR, UV-vis, XAS) to show a direct impact on the protonation equilibria, copper binding affinities, reduction potentials, and nitritereductase activities of these copper-peptide complexes. The potentials at a specific pH vary by 100 mV, and the nitritereductase activities range over a factor of 4 in rates. We also observe that the affinities, potentials, and catalytic activities are strongly influenced by the pH conditions (pH 5.8-7.4). In general, Cu(II) affinities for the peptides are diminished at low pH values. The interplay among these factors can lead to a 200 mV shift in reduction potential across these peptides, which is determined by the pH-dependent affinities of copper in both oxidation states. This study illustrates the strength of de novo protein design in elucidating the influence of ionizable residues on a particular redox system, an important step toward understanding the factors that govern the properties of this metalloenzyme with a goal of eventually improving the

Pseudoazurin (Paz) is the physiological electron donor to copper-containing nitritereductase (Nir), which catalyzes the reduction of NO2 (-) to NO. The Nir reaction mechanism involves the reduction of the type 1 (T1) copper electron transfer center by the external physiological electron donor, intramolecular electron transfer from the T1 copper center to the T2 copper center, and nitrite reduction at the type 2 (T2) copper catalytic center. We report the cloning, expression, and characterization of Paz from Sinorhizobium meliloti 2011 (SmPaz), the ability of SmPaz to act as an electron donor partner of S. meliloti 2011 Nir (SmNir), and the redox properties of the metal centers involved in the electron transfer chain. Gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis together with UV-vis and EPR spectroscopies revealed that as-purified SmPaz is a mononuclear copper-containing protein that has a T1 copper site in a highly distorted tetrahedral geometry. The SmPaz/SmNir interaction investigated electrochemically showed that SmPaz serves as an efficient electron donor to SmNir. The formal reduction potentials of the T1 copper center in SmPaz and the T1 and T2 copper centers in SmNir, evaluated by cyclic voltammetry and by UV-vis- and EPR-mediated potentiometric titrations, are against an efficient Paz T1 center to Nir T1 center to Nir T2 center electron transfer. EPR experiments proved that as a result of the SmPaz/SmNir interaction in the presence of nitrite, the order of the reduction potentials of SmNir reversed, in line with T1 center to T2 center electron transfer being thermodynamically more favorable.

The new copper(I) nitro complex [(Ph(3)P)(2)N][Cu(HB(3,5-Me(2)Pz)(3))(NO(2))] (2), containing the anionic hydrotris(3,5-dimethylpyrazolyl)borate ligand, was synthesized, and its structural features were probed using X-ray crystallography. Complex 2 was found to cocrystallize with a water molecule, and X-ray crystallographic analysis showed that the resulting molecule had the structure [(Ph(3)P)(2)N][Cu(HB(3,5-Me(2)Pz)(3))(NO(2))]·H(2)O (3), containing a water hydrogen bonded to an oxygen of the nitrite moiety. This complex represents the first example in the solid state of an analogue of the nitrous acid intermediate (CuNO(2)H). A comparison of the nitrite reduction reactivity of the electron-rich ligand containing the CuNO(2) complex 2 with that of the known neutral ligand containing the CuNO(2) complex [Cu(HC(3,5-Me(2)Pz)(3))(NO(2))] (1) shows that reactivity is significantly influenced by the electron density around the copper and nitrite centers. The detailed mechanisms of nitrite reduction reactions of 1 and 2 with acetic acid were explored by using density functional theory calculations. Overall, the results of this effort show that synthetic models, based on neutral HC(3,5-Me(2)Pz)(3) and anionic [HB(3,5-Me(2)Pz)(3)](-) ligands, mimic the electronic influence of (His)(3) ligands in the environment of the type II copper center of coppernitritereductases (Cu-NIRs).

Nitrite is converted to nitric oxide by haem or copper-containing enzymes in denitrifying bacteria during the process of denitrification. In designing an efficient biosensor, this enzymic turnover must be quantitatively assessed. The enzyme nitritereductase from Alcaligenes faecalis contains a redo

The crystal structures of copper-containing nitritereductase (CuNiR) from the thermophilic Gram-positive bacterium Geobacillus kaustophilus HTA426 and the amino (N)-terminal 68 residue-deleted mutant were determined at resolutions of 1.3Å and 1.8Å, respectively. Both structures show a striking resemblance with the overall structure of the well-known CuNiRs composed of two Greek key β-barrel domains; however, a remarkable structural difference was found in the N-terminal region. The unique region has one β-strand and one α-helix extended to the northern surface of the type-1 copper site. The superposition of the Geobacillus CuNiR model on the electron-transfer complex structure of CuNiR with the redox partner cytochrome c551 in other denitrifier system led us to infer that this region contributes to the transient binding with the partner protein during the interprotein electron transfer reaction in the Geobacillus system. Furthermore, electron-transfer kinetics experiments using N-terminal residue-deleted mutant and the redox partner, Geobacillus cytochrome c551, were carried out. These structural and kinetics studies demonstrate that the region is directly involved in the specific partner recognition.

Copper complexes of the deprotonated tridentate ligand, N-2-methylthiophenyl-2'-pyridinecarboxamide (HL1), were synthesized and characterized as part of our investigation into the reduction of copper(II) o-nitrito complexes into the related copper nitric oxide complexes and subsequent evolution of NO(g) such as occurs in the enzyme coppernitritereductase. Our studies afforded the complexes [(L1)Cu(II)Cl]n (1), [(L1)Cu(II)(ONO)] (2), [(L1)Cu(II)(H2O)](ClO4)·H2O (3·H2O), [(L1)Cu(II)(CH3OH)](ClO4) (4), [(L1)Cu(II)(CH3CO2)]·H2O (5·H2O), and [Co(Cp)2][(L1)Cu(I)(NO2)(CH3CN)] (6). X-ray crystal structure determinations revealed distorted square-pyramidal coordination geometry around Cu(II) ion in 1-5. Substitution of the H2O of 3 by nitrite quantitatively forms 2, featuring the κ(2)-O,O binding mode of NO2(-) to Cu(II). Reduction of 2 generates two Cu(I) species, one with κ(1)-O and other with the κ(1)-N bonded NO2(-) group. The Cu(I) analogue of 2, compound 6, was synthesized. The FTIR spectrum of 6 reveals the presence of κ(1)-N bonded NO2(-). Constant potential electrolysis corresponding to Cu(II) → Cu(I) reduction of a CH3CN solution of 2 followed by reaction with acids, CH3CO2H or HClO4 generates 5 or 3, and NO(g), identified electrochemically. The isolated Cu(I) complex 6 independently evolves one equivalent of NO(g) upon reaction with acids. Production of NO(g) was confirmed by forming [Co(TPP)NO] in CH2Cl2 (λ(max) in CH2Cl2: 414 and 536 nm, ν(NO) = 1693 cm(-1)).

Nitrate assimilation in many plants, algae, yeasts and bacteria is mediated by two enzymes, nitrate reductase (EC 1.6.6.2) and nitritereductase (EC 1.7.7.1). They catalyse the stepwise reduction of nitrate to nitrite and nitrite to ammonia respectively. The nitritereductase from an industrially important yeast, Candida utilis, has been purified to homogeneity. Purified nitritereductase is a heterodimer and the molecular masses of the two subunits are 58 and 66 kDa. The native enzyme exhibits a molecular mass of 126 kDa as analysed by gel filtration. The identify of the two subunits of nitritereductase was confirmed by immunoblotting using antibody for Cucurbita pepo leaf nitritereductase. The presence of two different sized transcripts coding for the two subunits was confirmed by (a) in vitro translation of mRNA from nitrate-induced C. utilis followed by immunoprecipitation of the in vitro translated products with heterologous nitritereductase antibody and (b) Northern-blot analysis. The 66 kDa subunit is acidic in nature which is probably due to its phosphorylated status. The enzyme is stable over a range of temperatures. Both subunits can catalyse nitrite reduction, and the reconstituted enzyme, at a higher protein concentration, shows an activity similar to that of the purified enzyme. Each of these subunits has been shown to contain a few unique peptides in addition to a large number of common peptides. Reduced Methyl Viologen has been found to be as effective an electron donor as NADPH in the catalytic process, a phenomenon not commonly seen for nitritereductases from other systems.

Nitritereductase (NiR) is a multicopper protein, with a trimeric structure containing two types of copper site: type I is present in each subunit whereas type 2 is localized at the subunits interface. The paper reports on the thermal behaviour of wild type NiR from Alcaligenes faecalis S-6. The

nitritereductase has been determined in both fully oxidized and reduced states. Intramolecular electron transfer (ET), between c and d(1) hemes is an essential step in the catalytic cycle. In earlier studies of the Pseudomonas stutzeri enzyme, we observed that a marked negative cooperativity......The cd(1) nitritereductases, which catalyze the reduction of nitrite to nitric oxide, are homodimers of 60 kDa subunits, each containing one heme-c and one heme-d(1). Heme-c is the electron entry site, whereas heme-d(1) constitutes the catalytic center. The 3D structure of Pseudomonas aeruginosa......-controlled bimolecular process, followed by unimolecular electron equilibration between the c and d(1) hemes (k(ET) = 4.3 s(-1) and K = 1.4 at 298 K, pH 7.0). In the case of the mutant, the latter ET rate was faster by almost one order of magnitude. Moreover, the internal ET rate dropped (by approximately 30-fold...

Full Text Available Nitrite and nitrate restore deficient endogenous nitric oxide (NO production as they are converted back to NO, and therefore complement the classic enzymatic NO synthesis. Circulating nitrate and nitrite must cross membrane barriers to produce their effects and increased nitrate concentrations may attenuate the nitrite influx into cells, decreasing NO generation from nitrite. Moreover, xanthine oxidoreductase (XOR mediates NO formation from nitrite and nitrate. However, no study has examined whether nitrate attenuates XOR-mediated NO generation from nitrite. We hypothesized that nitrate attenuates the vascular and blood pressure responses to nitrite either by interfering with nitrite influx into vascular tissue, or by competing with nitrite for XOR, thus inhibiting XOR-mediated NO generation. We used two independent vascular function assays in rats (aortic ring preparations and isolated mesenteric arterial bed perfusion to examine the effects of sodium nitrate on the concentration-dependent responses to sodium nitrite. Both assays showed that nitrate attenuated the vascular responses to nitrite. Conversely, the aortic responses to the NO donor DETANONOate were not affected by sodium nitrate. Further confirming these results, we found that nitrate attenuated the acute blood pressure lowering effects of increasing doses of nitrite infused intravenously in freely moving rats. The possibility that nitrate could compete with nitrite and decrease nitrite influx into cells was tested by measuring the accumulation of nitrogen-15-labeled nitrite (15N-nitrite by aortic rings using ultra-performance liquid chromatography tandem mass-spectrometry (UPLC-MS/MS. Nitrate exerted no effect on aortic accumulation of 15N-nitrite. Next, we used chemiluminescence-based NO detection to examine whether nitrate attenuates XOR-mediated nitritereductase activity. Nitrate significantly shifted the Michaelis Menten saturation curve to the right, with a 3-fold increase in

A conductometric biosensor for nitrite detection was developed using cytochrome c nitritereductase (ccNiR) extracted from Desulfovibrio desulfuricans ATCC 27774 cells immobilized on a planar interdigitated electrode by cross-linking with saturated glutaraldehyde (GA) vapour in the presence of bovine serum albumin, methyl viologen (MV), Nafion, and glycerol. The configuration parameters for this biosensor, including the enzyme concentration, ccNiR/BSA ratio, MV concentration, and Nafion concentration, were optimized. Various experimental parameters, such as sodium dithionite added, working buffer solution, and temperature, were investigated with regard to their effect on the conductance response of the biosensor to nitrite. Under the optimum conditions at room temperature (about 25 degrees C), the conductometric biosensor showed a fast response to nitrite (about 10s) with a linear range of 0.2-120 microM, a sensitivity of 0.194 microS/microM [NO(2)(-)], and a detection limit of 0.05 microM. The biosensor also showed satisfactory reproducibility (relative standard deviation of 6%, n=5). The apparent Michaelis-Menten constant (K(M,app)) was 338 microM. When stored in potassium phosphate buffer (100mM, pH 7.6) at 4 degrees C, the biosensor showed good stability over 1 month. No obvious interference from other ionic species familiar in natural waters was detected. The application experiments show that the biosensor is suitable for use in real water samples.

The enzyme nitritereductase (EC 1.6.6.4) is generally assayed in terms of disappearance of nitrite from the assay medium. We describe a technique which allowed estimation of the enzyme level in leaf tissues of Vigna mungo (L). Hepper in terms of the release of the product (NH3) of the enzyme reaction. The technique is offered as an alternative, possibly more convenient method for assay of nitritereductase in plant tissue in vivo.

Proton-coupled electron transfer (PCET), a ubiquitous phenomenon in biological systems, plays an essential role in coppernitritereductase (CuNiR), the key metalloenzyme in microbial denitrification of the global nitrogen cycle. Analyses of the nitrite reduction mechanism in CuNiR with conventional synchrotron radiation crystallography (SRX) have been faced with difficulties, because X-ray photoreduction changes the native structures of metal centers and the enzyme-substrate complex. Using serial femtosecond crystallography (SFX), we determined the intact structures of CuNiR in the resting state and the nitrite complex (NC) state at 2.03- and 1.60-Å resolution, respectively. Furthermore, the SRX NC structure representing a transient state in the catalytic cycle was determined at 1.30-Å resolution. Comparison between SRX and SFX structures revealed that photoreduction changes the coordination manner of the substrate and that catalytically important His255 can switch hydrogen bond partners between the backbone carbonyl oxygen of nearby Glu279 and the side-chain hydroxyl group of Thr280. These findings, which SRX has failed to uncover, propose a redox-coupled proton switch for PCET. This concept can explain how proton transfer to the substrate is involved in intramolecular electron transfer and why substrate binding accelerates PCET. Our study demonstrates the potential of SFX as a powerful tool to study redox processes in metalloenzymes.

Variants of the copper-containing nitritereductase (NiR) of Alcaligenes faecalis S6 were constructed by site-directed mutagenesis, by which the C-terminal histidine ligand (His145) of the Cu in the type-1 site was replaced by an alanine or a glycine. The type-1 sites in the NiR variants as isolated

Cytochrome cd1 nitritereductase is a bifunctional multiheme enzyme catalyzing the one-electron reduction of nitrite to nitric oxide and the four-electron reduction of dioxygen to water. Kinetics and thermodynamics of the internal electron transfer process in the Pseudomonas stutzeri enzyme have...

Objective To develtop directly molecular evolution of nitrite oxido-reductase using DNA-shuffling technique because nitrobacteria grow extremely slow and are unable to nitrify effectively inorganic nitrogen in wastewater treatment. Methods The norB gene coding the nitrite oxido-reductase in nitrobacteria was cloned and sequenced. Then, directed molecular evolution of nitrite oxido-reductase was developed by DNA-shuffling of 15 norB genes from different nitrobacteria. Results After DNA-shuffling with sexual PCR and staggered extension process PCR, the sequence was different from its parental DNA fragments and the homology ranged from 98% to 99%. The maximum nitrification rate of the modified bacterium of X16 by modified bacterium had the same characteristics of its parental bacteria of E. coli and could grow rapidly in normal cultures.Conclusion DNA-shuffling was successfully used to engineer E. coli, which had norB gene and could degrade inorganic nitrogen effectively.

The interaction of copper-containing dissimilatory nitritereductase from Alcaligenes faecalis S-6 ( AfNiR) with each of five small molecules was studied using crystallography and steady-state kinetics. Structural studies revealed that each small molecule interacted with the oxidized catalytic type 2 copper of AfNiR. Three small molecules (formate, acetate and nitrate) mimic the substrate by having at least two oxygen atoms for bidentate coordination to the type 2 copper atom. These three anions bound to the copper ion in the same asymmetric, bidentate manner as nitrite. Consistent with their weak inhibition of the enzyme ( K i >50 mM), the Cu-O distances in these AfNiR-inhibitor complexes were approximately 0.15 A longer than that observed in the AfNiR-nitrite complex. The binding mode of each inhibitor is determined in part by steric interactions with the side chain of active site residue Ile257. Moreover, the side chain of Asp98, a conserved residue that hydrogen bonds to type 2 copper-bound nitrite and nitric oxide, was either disordered or pointed away from the inhibitors. Acetate and formate inhibited AfNiR in a mixed fashion, consistent with the occurrence of second acetate binding site in the AfNiR-acetate complex that occludes access to the type 2 copper. A fourth small molecule, nitrous oxide, bound to the oxidized metal in a side-on fashion reminiscent of nitric oxide to the reduced copper. Nevertheless, nitrous oxide bound at a farther distance from the metal. The fifth small molecule, azide, inhibited the reduction of nitrite by AfNiR most strongly ( K ic = 2.0 +/- 0.1 mM). This ligand bound to the type 2 copper center end-on with a Cu-N c distance of approximately 2 A, and was the only inhibitor to form a hydrogen bond with Asp98. Overall, the data substantiate the roles of Asp98 and Ile257 in discriminating substrate from other small anions.

, internal electron transfer between these sites is an inherent element in the catalytic cycle of this enzyme. We have investigated the internal electron transfer reaction employing pulse radiolytically produced N-methyl nicotinamide radicals as reductant which reacts solely with the heme-c in an essentially...... diffusion controlled process. Following this initial step, the reduction equivalent is equilibrating between the c and d(1) heme sites in a unimolecular process (k=23 s(-1), 298 K, pH 7.0) and an equilibrium constant of 1.0. The temperature dependence of this internal electron transfer process has been......Cytochrome cd(1) nitritereductase from Pseudomonas stutzeri catalyzes the one electron reduction of nitrite to nitric oxide. It is a homodimer, each monomer containing one heme-c and one heme-d(1), the former being the electron uptake site while the latter is the nitrite reduction site. Hence...

Full Text Available Inclusion into the medium of 5 mg•dm-3 of non-ionic (ENF or ionic (DBST surfactant caused 50-60% inhibition of nitritereductase MR activity in S. polyrrhiza. At the same time, increased accumulation of NO2- in the plant tissues and lowering of the total and soluble protein contents were found. DBST also lowered the nitrate reductase (NR activity and the dry mass of the plants.

Full Text Available Background: Transfusion of blood remains the gold standard for fluid resuscitation from hemorrhagic shock. Hemoglobin (Hb within the red blood cell transports oxygen and modulates nitric oxide (NO through NO scavenging and nitritereductase. Aims: This study was designed to examine the effects of incorporating a novel NO modulator, RRx-001, on systemic and microvascular hemodynamic response after blood transfusion for resuscitation from hemorrhagic shock in a hamster window chamber model. In addition, to RRx-001 the role of low dose of nitrite (1 × 10−9 moles per animal supplementation after resuscitation was studied. Materials and Methods: Severe hemorrhage was induced by arterial controlled bleeding of 50% of the blood volume (BV and the hypovolemic state was maintained for 1 h. The animals received volume resuscitation by an infusion of 25% of BV using fresh blood alone or with added nitrite, or fresh blood treated with RRx-001 (140 mg/kg or RRx-001 (140 mg/kg with added nitrite. Systemic and microvascular hemodynamics were followed at baseline and at different time points during the entire study. Tissue apoptosis and necrosis were measured 8 h after resuscitation to correlate hemodynamic changes with tissue viability. Results: Compared to resuscitation with blood alone, blood treated with RRx-001 decreased vascular resistance, increased blood flow and functional capillary density immediately after resuscitation and preserved tissue viability. Furthermore, in RRx-001 treated animals, both mean arterial pressure (MAP and met Hb were maintained within normal levels after resuscitation (MAP >90 mmHg and metHb <2%. The addition of nitrite to RRx-001 did not significantly improve the effects of RRx-001, as it increased methemoglobinemia and lower MAP. Conclusion: RRx-001 alone enhanced perfusion and reduced tissue damage as compared to blood; it may serve as an adjunct therapy to the current gold standard treatment for resuscitation from

1. 1. The influence of copper on purified human erythrocyte glutathione reductase (E.C. 1.6.4.2) was studied. The holoenzyme was inhibited at low oxidized glutathione (GSSG) concentrations. At a glutathione concentration of 1 mM and higher no inhibition at all was found. The inhibition was independe

The reduction of nitrite into nitric oxide (NO) in denitrifying bacteria is catalyzed by nitritereductase. In several species, this enzyme is a heme-containing protein with one c heme and one d1 heme per monomer (cd1NiR), encoded by the nirS gene. For many years, the evidence of a link between NO and this hemeprotein represented a paradox, given that NO was known to tightly bind and, possibly, inhibit hemeproteins, including cd1NiRs. It is now established that, during catalysis, cd1NiRs diverge from "canonical" hemeproteins, since the product NO rapidly dissociates from the ferrous d1 heme, which, in turn, displays a peculiar "low" affinity for NO (KD=0.11 μM at pH 7.0). It has been also previously shown that the c heme reacts with NO at acidic pH but c heme nitrosylation was not extensively investigated, given that in cd1NiR it was considered a side reaction, rather than a genuine process controlling catalysis. The spectroscopic study of the reaction of cd1NiR and its semi-apo derivative (containing the sole c heme) with NO reported here shows that c heme nitrosylation is enhanced during catalysis; this evidence has been discussed in order to assess the potential of c heme nitrosylation as a regulatory process, as observed for cytochrome c nitrosylation in mammalian mitochondria.

Three Staphylococcus strains, S. carnosus, S. simulans and S. saprophyticus, selected due to their varying nitrite and/or nitrate-reductase activities, were used to initiate colour formation during sausage fermentation. During fermentation of sausages with either nitrite or nitrate added, colour...... nitrate depended on the specific Staphylococcus strain. Strains with high nitrate-reductase activity showed a significantly faster rate of pigment formation, but other factors were of influence as well. Product stability for the sliced, packaged sausage was evaluated as surface colour and oxidation...... by autofluorescence and hexanal content, respectively. No significant direct effect of the Staphylococcus addition was observed, however, there was a clear correspondence between high initial amount of MbFeIINO in the different sausages and the colour stability during storage. Autofluorescence data correlated well...

Sulphate-reducing bacteria (SRB) can be inhibited by nitrate-reducing, sulphide-oxidizing bacteria (NR-SOB), despite the fact that these two groups are interdependent in many anaerobic environments. Practical applications of this inhibition include the reduction of sulphide concentrations in oil fields by nitrate injection. The NR-SOB Thiomicrospira sp. strain CVO was found to oxidize up to 15 mM sulphide, considerably more than three other NR-SOB strains that were tested. Sulphide oxidation increased the environmental redox potential (Eh) from -400 to +100 mV and gave 0.6 nitrite per nitrate reduced. Within the genus Desulfovibrio, strains Lac3 and Lac6 were inhibited by strain CVO and nitrate for the duration of the experiment, whereas inhibition of strains Lac15 and D. vulgaris Hildenborough was transient. The latter had very high nitritereductase (Nrf) activity. Southern blotting with D. vulgaris nrf genes as a probe indicated the absence of homologous nrf genes from strains Lac3 and Lac6 and their presence in strain Lac15. With respect to SRB from other genera, inhibition of the known nitrite reducer Desulfobulbus propionicus by strain CVO and nitrate was transient, whereas inhibition of Desulfobacterium autotrophicum and Desulfobacter postgatei was long-lasting. The results indicate that inhibition of SRB by NR-SOB is caused by nitrite production. Nrf-containing SRB can overcome this inhibition by further reducing nitrite to ammonia, preventing a stalling of the favourable metabolic interactions between these two bacterial groups. Nrf, which is widely distributed in SRB, can thus be regarded as a resistance factor that prevents the inhibition of dissimilatory sulphate reduction by nitrite.

The objective of this study was to examine the expression and activity of nitrate reductase (NR, EC 1.7.1.1), nitritereductase (NiR, EC 1.7.2.2), glutamine synthetase (GS, EC 6.3.1.2), and glutamate synthase (GOGAT, EC 1.4.7.1) in response to potassium nitrate, ammonium chloride, and ammonium nitrate in nitrogen-starved wheat seedlings. Plants were grown in standard nutrient solution for 17 days and then subjected to nitrogen starvation for 7 days. The starved plants were supplied with potassium nitrate ammonium nitrate and ammonium chloride (50 mM) for 4 days and the leaves were harvested. The relative expression of NR, NiR, GS, and GOGAT as well as the enzyme activities were investigated. Nitrogen starvation caused a significant decrease both in transcript levels and in NR, NiR, GS, and GOGAT activities. Potassium nitrate and ammonium nitrate treatments restored NR, NiR, GS, and GOGAT expressions and activities. Ammonium chloride increased only the expressions and activities of GS and GOGAT in a dose-dependent manner. The results of our study highlight the differential effects between the type and the amount of nitrogen salts on NR, NiR, GS, and GOGAT activities in wheat seedlings while potassium nitrate being more effective.

Nanostructured polypyrrole film was synthesized onto a copper electrode in solutions of oxalic and salicylic acids and their buffers. The electrooxidation of pyrrole to form polypyrrole film and the electroreduction of nitrate and nitrite ions at synthesized Ppy modified copper electrodes (Ppy/Cu) in potassium chloride aqueous solutions were studied using chronoamperometry. The nanoporous structure of the synthesized Ppy films was characterized by scanning electron microscopy (SEM). Nitrate and nitrite reduction were performed by an electrochemical method under potentiostatic conditions. The Ppy/Cu electrodes prepared in the oxalate buffer and salicylic acid solutions perform more stable catalytic activity for nitrate reduction; their service life is about ten times longer than that for the electrodes prepared in oxalic acid solution. After 20 h of electrolysis, the nitrite was reduced completely with 100% efficiency and the nitrate was reduced with 35% efficiency. Report submitted to the 5th International Workshop on Advanced Materials Science and Nanotechnology IWAMSN, Hanoi, 9-12 November 2010.

The genetic heterogeneity of nitritereductase gene (nirK and nirS) fragments from denitrifying prokaryotes in forested upland and marsh soil was investigated using molecular methods. nirK gene fragments could be amplified from both soils, whereas nirS gene fragments could be amplified only from...... the marsh soil. PCR products were cloned and screened by restriction fragment length polymorphism (RFLP), and representative fragments were sequenced. The diversity of nirK clones was lower than the diversity of nirS clones. Among the 54 distinct nirK RFLP patterns identified in the two soils, only one...... marsh clones and all upland clones. Only a few of the nirK clone sequences branched with those of known denitrifying bacteria. The nirS clones formed two major clusters with several subclusters, but all nirS clones showed less than 80% identity to nirS sequences from known denitrifying bacteria. Overall...

The high-yield expression and purification of Shewanella oneidensis cytochrome c nitritereductase (ccNiR) and its characterization by a variety of methods, notably Laue crystallography, are reported. A key component of the expression system is an artificial ccNiR gene in which the N-terminal signal peptide from the highly expressed S. oneidensis protein 'small tetraheme c' replaces the wild-type signal peptide. This gene, inserted into the plasmid pHSG298 and expressed in S. oneidensis TSP-1 strain, generated approximately 20 mg crude ccNiR per liter of culture, compared with 0.5-1 mg/L for untransformed cells. Purified ccNiR has nitrite and hydroxylamine reductase activities comparable to those previously reported for Escherichia coli ccNiR, and is stable for over 2 weeks in pH 7 solution at 4 C. UV/vis spectropotentiometric titrations and protein film voltammetry identified five independent one-electron reduction processes. Global analysis of the spectropotentiometric data also allowed determination of the extinction coefficient spectra for the five reduced ccNiR species. The characteristics of the individual extinction coefficient spectra suggest that, within each reduced species, the electrons are distributed among the various hemes, rather than being localized on specific heme centers. The purified ccNiR yielded good-quality crystals, with which the 2.59-{angstrom}-resolution structure was solved at room temperature using the Laue diffraction method. The structure is similar to that of E. coli ccNiR, except in the region where the enzyme interacts with its physiological electron donor (CymA in the case of S. oneidensis ccNiR, NrfB in the case of the E. coli protein).

The phytochrome-deficient aurea mutant of tomato (Lycopersicon esculentum (L.) Mill) was used to investigate if phytochrome plays a role in the regulation of nitrate-reductase (NR, EC 1.6.6.1) and nitrite-reductase (NiR, EC 1.7.7.1) gene expression. We show that the expression of the tomato NR and NiR genes is stimulated by light and that this light response is mediated by the photoreceptor phytochrome. The red-light response of the NR and NiR genes was reduced in etiolated aurea seedlings when compared to isogenic wild-type cotyledons. The relative levels of NR mRNA and NiR transcripts and their diurnal fluctuations were identical in mature white-light-grown leaves of the wild-type and of the aurea mutant. The transcript levels for cab and RbcS (genes for the chlorophyll-a/b-binding protein of PSII and the small subunit of the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase, respectively) in aurea leaves grown in white light were indistinguishable from the respective transcript levels in the leaves of the wildtype grown under the same conditions. Despite a severe reduction in the chlorophyll content, the rate of net CO2 uptake by leaves of the aurea mutant was only slightly reduced when compared to the rate of net photosynthesis of wild-type leaves. This difference in the photosynthetic performances of wild-type and aurea mutant plants disappeared during aging of the plants. The increase in zeaxanthin and the concomitant decrease in violaxanthin in leaves of the aurea mutant compared with the same pigment levels in leaves of the wild-type indicate that the activity of the xanthophyll cycle is increased in aurea leaves as a consequence of the reduced CO2-fixation capacity of the mutant leaves.

A rapid and efficient copper-catalyzed nitration of aryl halides has been established under microwave irradiation. The catalytic systems were found to be the most effective with 4-substituted aryl iodides leading to nearly complete conversions. Nitration of aromatic compounds is one of the important industrial processes as underlying intermediates in the manufacture of a wide range of chemicals such as dyes, pharmaceuticals, agrochemicals and explosives. General methods for the nitration of aromatic compounds utilize strongly acidic conditions employing nitric acid or a mixture of nitric and sulfuric acids, sometimes leading to problems with poor regioselectivity, overnitration, oxidized byproducts and excess acid waste in many cases of functionalized aromatic compounds. Several other nitrating agents or methods avoiding harsh reaction conditions have been explored using metal nitrates, nitrite salts, and ionic liquid-mediated or microwave-assisted nitrations. Recently, copper or palladium compounds have been successfully used as efficient catalysts for the arylation of amines with aryl halides under mild conditions.

Copper-cobalt bimetal nanoparticles (Cu-Co) have been electrochemically prepared on glassy carbon electrodes (GCEs), which were electrodeposited with conducting polymer nanocomposites of poly(3,4-ethylenedioxythiophene) (PEDOT) doped with carbon nanotubes (CNTs). Owing to their good conductivity, high mechanical strength, and large surface area, the PEDOT/CNTs composites offered excellent substrates for the electrochemical deposition of Cu-Co nanoparticles. As a result of their nanostructure and the synergic effect between Cu and Co, the Cu-Co/PEDOT/CNTs composites exhibited significantly enhanced catalytic activity towards the electrochemical oxidation of nitrite. Under optimized conditions, the nanocomposite-modified electrodes had a fast response time within 2 s and a linear range from 0.5 to 430 μm for the detection of nitrite, with a detection limit of 60 nm. Moreover, the Cu-Co/PEDOT/CNTs composites were highly stable, and the prepared nitrite sensors could retain more than 96 % of their initial response after 30 days.

Escherichia coli synthesize C-type cytochromes only during anaerobic growth in media supplemented with nitrate and nitrite. The reduction of nitrate to ammonium in the periplasm of Escherichia coli involves two separate periplasmic enzymes, nitrate reductase and nitritereductase. The nitritereductase involved, NrfA, contains cytochrome C and is synthesized coordinately with a membrane-associated cytochrome C, NrfB, during growth in the presence of nitrite or in limiting nitrate concentrations. The genes NrfE, NrfF, and NrfG are required for the formate-dependent nitrite reduction pathway, which involves at least two C-type cytochrome proteins, NrfA and NrfB. The NrfE, NrfF, and NrfG genes (heme lyase complex) are involved in the maturation of a special C-type cytochrome, apocytochrome C (apoNrfA), to cytochrome C (NrfA) by transferring a heme to the unusual heme binding motif of the Cys-Trp-Ser-Cys-Lys sequence in apoNrfA protein. Thus, in order to further investigate the roles of NrfG in the formation of heme lyase complex (NrfEFG) and in the interaction between heme lyase complex and formate-dependent nitritereductase (NrfA), we determined the crystal structure of NrfG at 2.05 A. The structure of NrfG showed that the contact between heme lyase complex (NrfEFG) and NrfA is accomplished via a TPR domain in NrfG which serves as a binding site for the C-terminal motif of NrfA. The portion of NrfA that binds to TPR domain of NrfG has a unique secondary motif, a helix followed by about a six-residue C-terminal loop (the so called "hook conformation"). This study allows us to better understand the mechanism of special C-type cytochrome assembly during the maturation of formate-dependent nitritereductase, and also adds a new TPR binding conformation to the list of TPR-mediated protein-protein interactions.

Full Text Available Abstract Background The detection of hydrogen peroxide (H2O2 and nitrite ion (NO2- is of great important in various fields including clinic, food, pharmaceutical and environmental analyses. Compared with many methods that have been developed for the determination of them, the electrochemical detection method has attracted much attention. In recent years, with the development of nanotechnology, many kinds of micro/nano-scale materials have been used in the construction of electrochemical biosensors because of their unique and particular properties. Among these catalysts, copper oxide (CuO, as a well known p-type semiconductor, has gained increasing attention not only for its unique properties but also for its applications in many fields such as gas sensors, photocatalyst and electrochemistry sensors. Continuing our previous investigations on transition-metal oxide including cuprous oxide and α-Fe2O3 modified electrode, in the present paper we examine the electrochemical and electrocatalytical behavior of flower like copper oxide modified glass carbon electrodes (CuO/GCE. Results Flower like copper oxide (CuO composed of many nanoflake was synthesized by a simple hydrothermal reaction and characterized using field-emission scanning electron microscopy (FE-SEM and X-ray diffraction (XRD. CuO modified glass carbon electrode (CuO/GCE was fabricated and characterized electrochemically. A highly sensitive method for the rapid amperometric detection of hydrogen peroxide (H2O2 and nitrite (NO2- was reported. Conclusions Due to the large specific surface area and inner characteristic of the flower like CuO, the resulting electrode show excellent electrocatalytic reduction for H2O2 and oxidation of NO2-. Its sensitivity, low detection limit, fast response time and simplicity are satisfactory. Furthermore, this synthetic approach can also be applied for the synthesis of other inorganic oxides with improved performances and they can also be extended to

采用rpиcc试剂比色法,研究了VC对灰棕紫泥土亚硝酸还原酶动力学参数的影响.结果表明,VC的加入降低了V0、Km和Vmax值,表明VC对土壤亚硝酸还原酶的作用类型表现为反竞争性抑制作用,但低浓度VC会减弱亚硝酸还原酶与底物亲和力,而加快产物与酶的分离.由于VC具有抗氧化作用,对土壤具有良好作用,作为安全、有效的反硝化抑制剂使用.%The article researched the effect of Vc treatment on the nitritereductase kinetic parameter of gray-brown purple clay soil by rpHCC reagent colorimetric method. The result showed that Vc treatment reduced the values of V0, Km and Vmax, the effect type of Vc on soil nitritereductase was uncompetitive inhibition. Low concentration Vc reduced the affinity of nitritereductase with substrate, and accelerated the separation of products and enzyme. Vc has antioxidant effect, and is good for to soil, it could be the safe and functional nitrification inhibitors.

The cytochrome c nitritereductase NrfA is a 53 kDa pentahaem enzyme that crystallizes as a decahaem homodimer. NrfA catalyses the reduction of NO2- to NH4+ through a six electron reduction pathway that is of major physiological significance to the anaerobic metabolism of enteric and sulfate reducing bacteria. NrfA receives electrons from the 21 kDa pentahaem NrfB donor protein. This requires that redox complexes form between the NrfA and NrfB pentahaem cytochromes. The formation of these complexes can be monitored using a range of methodologies for studying protein-protein interactions, including dynamic light scattering, gel filtration, analytical ultracentrifugation and visible spectroscopy. These methods have been used to show that oxidized NrfA exists in dynamic monomer-dimer equilibrium with a Kd (dissociation constant) of 4 μM. Significantly, the monomeric and dimeric forms of NrfA are equally active for either the six electron reduction of NO2- or HSO3-. When mixed together, NrfA and NrfB exist in equilibrium with NrfAB, which is described by a Kd of 50 nM. Thus, since NrfA and NrfB are present in micromolar concentrations in the periplasmic compartment, it is likely that NrfB remains tightly associated with its NrfA redox partner under physiological conditions.

Nitrite is presently considered a NO "storage form" that can be made available, through its one-electron reduction, to maintain NO formation under hypoxia/anoxia. The molybdoenzymes xanthine oxidase/dehydrogenase (XO/XD) and aldehyde oxidase (AO) are two of the most promising mammalian nitritereductases, and in this work, we characterized NO formation by rat and human XO/XD and AO. This is the first characterization of human enzymes, and our results support the employment of rat liver enzymes as suitable models of the human counterparts. A comprehensive kinetic characterization of the effect of pH on XO and AO-catalyzed nitrite reduction showed that the enzyme's specificity constant for nitrite increase 8-fold, while the Km(NO2(-)) decrease 6-fold, when the pH decreases from 7.4 to 6.3. These results demonstrate that the ability of XO/AO to trigger NO formation would be greatly enhanced under the acidic conditions characteristic of ischemia. The dioxygen inhibition was quantified, and the Ki(O2) values found (24.3-48.8 μM) suggest that in vivo NO formation would be fine-tuned by dioxygen availability. The potential in vivo relative physiological relevance of XO/XD/AO-dependent pathways of NO formation was evaluated using HepG2 and HMEC cell lines subjected to hypoxia. NO formation by the cells was found to be pH-, nitrite-, and dioxygen-dependent, and the relative contribution of XO/XD plus AO was found to be as high as 50%. Collectively, our results supported the possibility that XO/XD and AO can contribute to NO generation under hypoxia inside a living human cell. Furthermore, the molecular mechanism of XO/AO-catalyzed nitrite reduction was revised.

Estuarine systems are the major conduits for the transfer of nitrate from agricultural and other terrestrial-anthropogenic sources into marine ecosystems. Within estuarine sediments some microbially driven processes (denitrification and anammox) result in the net removal of nitrogen from the environment, while others (dissimilatory nitrate reduction to ammonium) do not. In this study, molecular approaches have been used to investigate the diversity, abundance, and activity of the nitrate-reducing communities in sediments from the hypernutrified Colne estuary, United Kingdom, via analysis of nitrate and nitritereductase genes and transcripts. Sequence analysis of cloned PCR-amplified narG, napA, and nrfA gene sequences showed the indigenous nitrate-reducing communities to be both phylogenetically diverse and also divergent from previously characterized nitrate reduction sequences in soils and offshore marine sediments and from cultured nitrate reducers. In both the narG and nrfA libraries, the majority of clones (48% and 50%, respectively) were related to corresponding sequences from delta-proteobacteria. A suite of quantitative PCR primers and TaqMan probes was then developed to quantify phylotype-specific nitrate (narG and napA) and nitritereductase (nirS and nrfA) gene and transcript numbers in sediments from three sites along the estuarine nitrate gradient. In general, both nitrate and nitritereductase gene copy numbers were found to decline significantly (P < 0.05) from the estuary head towards the estuary mouth. The development and application, for the first time, of quantitative reverse transcription-PCR assays to quantify mRNA sequences in sediments revealed that transcript numbers for three of the five phylotypes quantified were greatest at the estuary head.

A system was developed for the detection of denitrifying bacteria by the application of specific nitritereductase gene fragments with PCR. Primer sequences were found for the amplification of fragments from both nitritereductase genes (nirK and nirS) after comparative sequence analysis. Whenever amplification was tried with these primers, the known nir type of denitrifying laboratory cultures could be confirmed. Likewise, the method allowed a determination of the nir type of five laboratory strains. The nirK gene could be amplified from Blastobacter denitrificans, Alcaligenes xylosoxidans, and Alcaligenes sp. (DSM 30128); the nirS gene was amplified from Alcaligenes eutrophus DSM 530 and from the denitrifying isolate IFAM 3698. For each of the two genes, at least one primer combination amplified successfully for all of the test strains. Specific amplification products were not obtained wit h nondenitrifying bacteria or with strains of the other nir type. The specificity of the amplified products was confirmed by subsequent sequencing. These results suggest the suitability of the method for the qualitative detection of denitrifying bacteria in environmental samples. This was shown by applying the generally amplifying primer combination for each nir gene developed in this study to total DNA preparations from aquatic habitats.

Nitrite reductase（NIR） reduces nitrite to NO or NH4＋,this is a key step in the nitrogen cycle.The advances in study on NIR,including the structure,catalysis mechanism and the response characteristic were reviewed.Application of NIR and denitrifying bacteria in meat products were summarized.Application and research prospects of NIR in the future were also presented.%亚硝酸还原酶（nitritereductase,NIR）是陆地上氮元素循环过程中降解亚硝酸盐的一个关键酶,能把亚硝酸盐还原生成NO或NH4＋。综述了国内外研究者从不同方面对NIR的研究,论述了NIR的组成结构、工作机理及反应特性,总结了产NIR菌株及NIR在肉制品中的应用,并对NIR的研究及应用进行了展望。

Full Text Available Cytochrome cd1 nitritereductases (cd1NiRs catalyze the one-electron reduction of nitrite to nitric oxide. Due to their catalytic reaction, cd1NiRs are regarded as promising components for biosensing, bioremediation and biotechnological applications. Motivated by earlier findings that catalytic activity of cd1NiR from Marinobacter hydrocarbonoclasticus (Mhcd1 depends on the presence of its physiological redox partner, cytochrome c552 (cyt c552, we show here a detailed surface enhanced resonance Raman characterization of Mhcd1 and cyt c552 attached to biocompatible electrodes in conditions which allow direct electron transfer between the conducting support and immobilized proteins. Mhcd1 and cyt c552 are co-immobilized on silver electrodes coated with self-assembled monolayers (SAMs and the electrocatalytic activity of Ag // SAM // Mhcd1 // cyt c552 and Ag // SAM // cyt c552 // Mhcd1 constructs is tested in the presence of nitrite. Simultaneous evaluation of structural and thermodynamic properties of the immobilized proteins reveals that cyt c552 retains its native properties, while the redox potential of apparently intact Mhcd1 undergoes a ~150 mV negative shift upon adsorption. Neither of the immobilization strategies results in an active Mhcd1, reinforcing the idea that subtle and very specific interactions between Mhcd1 and cyt c552 govern efficient intermolecular electron transfer and catalytic activity of Mhcd1.

A long-standing issue in protein film voltammetry (PFV), particularly electrocatalytic voltammetry of redox enzyme monolayers, is the variability of protein adsorption modes, reflected in distributions of catalytic activity of the adsorbed protein/enzyme molecules. Use of well-defined, atomically...

The increasing sequence information on oxygen reductases of the haem-copper superfamily, together with the available three-dimensional structures, allows a clear identification of their common, functionally important features. Taking into consideration both the overall amino acid sequences of the core subunits and key residues involved in proton transfer, a novel hypothesis for the molecular evolution of these enzymes is proposed. Three main families of oxygen reductases are identified on the basis of common features of the core subunits, constituting three lines of evolution: (i) type A (mitochondrial-like oxidases), (ii) type B (ba3-like oxidases) and (iii) type C (cbb3-type oxidases). The first group can be further divided into two subfamilies, according to the helix VI residues at the hydrophobic end of one of the proton pathways (the so-called D-channel): (i) type A1, comprising the enzymes with a glutamate residue in the motif -XGHPEV-, and (ii) type A2, enzymes having instead a tyrosine and a serine in the alternative motif -YSHPXV-. This second subfamily of oxidases is shown to be ancestor to the one containing the glutamate residue, which in the Bacteria domain is only present in oxidases from Gram-positive or purple bacteria. It is further proposed that the Archaea domain acquired terminal oxidases by gene transfer from the Gram-positive bacteria, implying that these enzymes were not present in the last common ancestor before the divergence between Archaea and Bacteria. In fact, most oxidases from archaea have a higher amino acid sequence identity and similarity with those from bacteria, mainly from the Gram-positive group, than with oxidases from other archaea. Finally, a possible relation between the dihaemic subunit (FixP) of the cbb3 oxidases and subunit II of caa3 oxidases is discussed. As the families of haem-copper oxidases can also be identified by their subunit II, a parallel evolution of subunits I and II is suggested.

The gene for cytochrome cd1 nitritereductase of Paracoccus pantotrophus, a protein of known crystal structure, is nirS. This gene is shown to be flanked by genes previously recognized in other organisms to encode proteins involved in the control of its transcription (nirI) and the biosynthesis of the d1 cofactor (nirE). Northern blot analysis has established under anaerobic conditions that a monocistronic transcript is produced from nirS, in contrast to observations with other denitrifying bacteria in which arrangement of flanking genes is different and the messages produced are polycistronic. The lack of a transcript under aerobic conditions argues against a role for cytochrome cd1 in the previously proposed aerobic denitrification pathway in Pa. pantotrophus. A putative rho-independent transcription termination sequence immediately following nirS, and preceding nirE, can be identified. The independent transcription of nirS and nirE indicates that it should be possible to produce site-directed mutants of nirS borne on a plasmid in a nirS deletion mutant. The transcript start point for nirS has been determined by two complementary techniques, 5'-RACE (Rapid amplification of cDNA 5' ends) and primer extension. It is 29 bp upstream of the AUG of nirS. An anaerobox, which presumably binds Nnr, is centred a further 41.5 bp upstream of the transcript start. No standard sigma70 DNA sequence motifs can be identified, but a conserved sequence (T-T-GIC-C-G/C-G/C) can be found in approximately the same position (-16) upstream of the transcript starts of nirS and nirI, whose products are both involved in the conversion of nitrite to nitric oxide.

[Objective]The study aimed to explore the mechanism of nitritereductase from Lactobacillus plantarum WU14 under nitrite stress so that lay a foundation for pure culture technology of lactic acid bacteria in fermented food. [Method] Growth density, pH and nitrite degradation quantity of L. plantarum WU14 were determined when the liquid medium contained sodium nitrite ranged from 0.02% to 0.16% under the condition of 37℃. The recombination strain Lactococcus lactic NZ9000/pRNA48-NirS was constructed followed the putative nitritereductase gene from L. plantarum WU14 was amplified by PCR and then cloned into the food-grade cytoplasmic inducible expression vector pRNA48 of L. lactic NZ9000. After induced with 30 ng·mL-1 nisin, the expressed target protein and the enzyme activity of nitritereductase of the recombinant strains were analyzed by SDS-PAGE and Naphthyl ethylenediamine dihydrochloride spectrophotometric method. Using the bioinformatics software, the high level protein structure, membrane structure and hydrophobicity of nitritereductase gene were predicted and analyzed. [Result]L. plantarum WU14 could routinely grow in MRS medium containing less than 0.12% nitrite, along with degradation of nitrite. After the strain L. plantarum WU14 was cultured for 24 hours in the liquid medium containing 0.10% sodium nitrite, the nitritereductase activity of L. plantarum WU14 was 2 347.5 U·mL-1, and the degradation quantity was 56.34 μg·mL-1 according to the analysis of its nitrite degradation ability. the NirS gene could express in the recombinant strain. Nitritereductase gene encodes a kind of hydrophilic protein containing alpha helix and random coil, no signal peptide and transmembrane structure. The recombination strain could routinely grow in GM17 medium containing less than 0.10% nitrite, meanwhile, its enzyme activity reached 925.41 U·mL-1 and the degradation quantity reached 22.21 mg·mL-1 after 24 h fermentation in the 0.04% nitrite concentration

The periplasmic cytochrome cd 1 nitritereductase NirS occurring in denitrifying bacteria such as the human pathogen Pseudomonas aeruginosa contains the essential tetrapyrrole cofactors haem c and haem d 1. Whereas the haem c is incorporated into NirS by the cytochrome c maturation system I, nothing is known about the insertion of the haem d 1 into NirS. Here, we show by co-immunoprecipitation that NirS interacts with the potential haem d 1 insertion protein NirN in vivo. This NirS–NirN inter...

Several spectroscopic methods, including absorption, circular dichroism (CD), magnetic CD (MCD), X-ray absorption, resonance Raman, EPR, NMR, and quasi-elastic light-scattering spectroscopy, have been used to probe the structures of copper-containing amine oxidases, nitritereductase, and nitrous oxide reductase. The basic goals are to determine the copper site structure, electronic properties, and to generate structure-reactivity correlations. Collectively, the results on the amine oxidases permit a detailed model for the Cu(II) sites in these enzymes to be constructed that, in turn, rationalizes the ligand-binding chemistry. Resonance Raman spectra of the phenylhydrazine and 2,4-dinitrophenyl-hydrazine derivatives of bovine plasma amine oxidase and models for its organic cofactor, e.g. pyridoxal, methoxatin, are most consistent with methoxatin being the intrinsic cofactor. The structure of the Cu(I) forms of the amine oxidases have been investigated by X-ray absorption spectroscopy (XAS); the copper coordination geometry is significantly different in the oxidized and reduced forms. Some anomalous properties of the amine oxidases in solution are explicable in terms of their reversible aggregation, which the authors have characterized via light scattering. Nitrite and nitrous oxide reductases display several novel spectral properties. The data suggest that new types of copper sites are present.

This report documents work at the U.S. Geological Survey (USGS) National Water Quality Laboratory (NWQL) to validate enzymatic reduction, colorimetric determinative methods for nitrate + nitrite in filtered water by automated discrete analysis. In these standard- and low-level methods (USGS I-2547-11 and I-2548-11), nitrate is reduced to nitrite with nontoxic, soluble nitrate reductase rather than toxic, granular, copperized cadmium used in the longstanding USGS automated continuous-flow analyzer methods I-2545-90 (NWQL laboratory code 1975) and I-2546-91 (NWQL laboratory code 1979). Colorimetric reagents used to determine resulting nitrite in aforementioned enzymatic- and cadmium-reduction methods are identical. The enzyme used in these discrete analyzer methods, designated AtNaR2 by its manufacturer, is produced by recombinant expression of the nitrate reductase gene from wall cress (Arabidopsis thaliana) in the yeast Pichia pastoris. Unlike other commercially available nitrate reductases we evaluated, AtNaR2 maintains high activity at 37°C and is not inhibited by high-phenolic-content humic acids at reaction temperatures in the range of 20°C to 37°C. These previously unrecognized AtNaR2 characteristics are essential for successful performance of discrete analyzer nitrate + nitrite assays (henceforth, DA-AtNaR2) described here.

以盐敏感荞麦品种TQ-0808和耐盐荞麦品种川荞1号为试验材料,采用NaCl和等渗PEG-6000处理,研究渗透胁迫和盐胁迫对不同耐盐性荞麦品种硝酸还原酶(NR)及亚硝酸还原酶(NiR)活性的影响.结果表明,高浓度盐胁迫下盐敏感荞麦品种叶片NR及NiR活性显著降低,而耐盐荞麦品种降低幅度相对较小,且高浓度盐胁迫下盐敏感荞麦品种叶片NR及NiR活性的降低幅度明显大于渗透胁迫的,说明Na+毒害效应发挥了主要作用.另外,两个荞麦品种叶片NR活性高低与其叶片硝酸盐含量呈正相关.%The salt-sensitive buckwheat variety (TQ-0808) and salt-tolerant buckwheat variety (Chuanqiao No.1)were used as experimental materials,which were treated with NaCl and iso-osmotic PEG-6000.The effects of osmotic and salt stress on nitrate reductase and nitritereductase activity of buckwheat varieties with different salt tolerance were studied.The results showed that the nitrate reductase and nitritereductase activity of leaves in salt-sensitive buckwheat variety were decreased significantly under the salt stress of high concentration,while that in salt-tolerant buckwheat variety were decreased relatively small,and that in salt-sensitive buckwheat variety under the salt stress of high concentration were obviously more than those under the osmotic stress.It indicated that ion toxicity effects of Na + play the major role.In addition,the level of nitrate reductase activity of leaves in two buckwheat varieties was positively correlated with the nitrate content of leaves.

We describe a simple electrochemical preparation method of a novel three dimensional (3D) graphene material, porous flower-like reduced graphene oxide (f-RGO) nanosheets, which was explored as the support for Cu particles on a glassy carbon electrode (Cu/f-RGO/GCE) for detecting nitrite. In morphology studies, scanning electron microscopy (SEM) demonstrates the 3D porous structure of f-RGO enlarges the surface area of the electrode and promotes more Cu particles depositing on the surface of f-RGO with homogeneous dispersion. In cyclic voltammetry (CV), a well-defined voltammetric peak along with the remarkable reduction current indicates excellent electrocatalytic activity of the Cu/f-RGO/GCE for NaNO2 reduction compared with other corresponding electrodes. The effects of pH value and detection potential on the current responses of Cu/f-RGO/GCE towards nitrite were optimized to obtain the maximal sensitivity. In the optimal experimental conditions, Cu/f-RGO/GCE displays the wide detection range from 0.15 μM to 10,500 μM and the low limit of detection of 0.06 μM (S/N = 3) with fast response time 2 s for detecting NaNO2 through an amperometric method. Furthermore, the presence of K(+), Na(+), Cl(-), NH4(+), NO3(-), SO4(2-) and ascorbic acid show a negligible effect on the current response of nitrite determination suggesting Cu/f-RGO/GCE have the high selectivity for detecting nitrite even in the presence of high concentration of interferents. Moreover, the real sample determination experiment indicated practical feasibility of the obtained sensor. The prepared sensor for determination of NaNO2 exhibited wide liner range, low detection limit, good reproducibility, nice stability and remarkable anti-interference ability. In this paper, not only did the Cu/f-RGO/GCE show high performance for determination of nitrite, but also it was simple to prepare, user-friendly and cost-effective.

Staphylococcus carnosus reduces nitrate to ammonia in two steps. (i) Nitrate was taken up and reduced to nitrite, and nitrite was subsequently excreted. (ii) After depletion of nitrate, the accumulated nitrite was imported and reduced to ammonia, which again accumulated in the medium. The localization, energy gain, and induction of the nitrate and nitritereductases in S. carnosus were characterized. Nitrate reductase seems to be a membrane-bound enzyme involved in respiratory energy conserva...

Staphylococcus carnosus reduces nitrate to ammonia in two steps. (i) Nitrate was taken up and reduced to nitrite, and nitrite was subsequently excreted. (ii) After depletion of nitrate, the accumulated nitrite was imported and reduced to ammonia, which again accumulated in the medium. The localization, energy gain, and induction of the nitrate and nitritereductases in S. carnosus were characterized. Nitrate reductase seems to be a membrane-bound enzyme involved in respiratory energy conservation, whereas nitritereductase seems to be a cytosolic enzyme involved in NADH reoxidation. Syntheses of both enzymes are inhibited by oxygen and induced to greater or lesser degrees by nitrate or nitrite, respectively. In whole cells, nitrite reduction is inhibited by nitrate and also by high concentrations of nitrite (> or = 10 mM). Nitrite did not influence nitrate reduction. Two possible mechanisms for the inhibition of nitrite reduction by nitrate that are not mutually exclusive are discussed. (i) Competition for NADH nitrate reductase is expected to oxidize the bulk of the NADH because of its higher specific activity. (ii) The high rate of nitrate reduction could lead to an internal accumulation of nitrite, possibly the result of a less efficient nitrite reduction or export. So far, we have no evidence for the presence of other dissimilatory or assimilatory nitrate or nitritereductases in S. carnosus.

A multiyear research effort at the U.S. Geological Survey (USGS) National Water Quality Laboratory (NWQL) evaluated several commercially available nitrate reductase (NaR) enzymes as replacements for toxic cadmium in longstanding automated colorimetric air-segmented continuous-flow analyzer (CFA) methods for determining nitrate plus nitrite (NOx) in water. This research culminated in USGS approved standard- and low-level enzymatic reduction, colorimetric automated discrete analyzer NOx methods that have been in routine operation at the NWQL since October 2011. The enzyme used in these methods (AtNaR2) is a product of recombinant expression of NaR from Arabidopsis thaliana (L.) Heynh. (mouseear cress) in the yeast Pichia pastoris. Because the scope of the validation report for these new automated discrete analyzer methods, published as U.S. Geological Survey Techniques and Methods 5–B8, was limited to performance benchmarks and operational details, extensive foundational research with different enzymes—primarily YNaR1, a product of recombinant expression of NaR from Pichia angusta in the yeast Pichia pastoris—remained unpublished until now. This report documents research and development at the NWQL that was foundational to development and validation of the discrete analyzer methods. It includes: (1) details of instrumentation used to acquire kinetics data for several NaR enzymes in the presence and absence of known or suspected inhibitors in relation to reaction temperature and reaction pH; and (2) validation results—method detection limits, precision and bias estimates, spike recoveries, and interference studies—for standard- and low-level automated colorimetric CFA-YNaR1 reduction NOx methods in relation to corresponding USGS approved CFA cadmium-reduction (CdR) NOx methods. The cornerstone of this validation is paired sample statistical and graphical analysis of NOx concentrations from more than 3,800 geographically and seasonally diverse surface

Methaemoglobinaemia is a potential toxic effect of aliphatic nitrites which are increasingly abused by male homosexuals and drug addicts because of marked vasodilating properties ('poppers'). In most instances, severe complications were described following the ingestion of large quantities of amyl, butyl or isobutyl nitrites. A deficiency in NADH-dependent haemoglobin reductase in some patients has been noted. This is the first report of symptomatic methaemoglobinaemia following the inhalation of amyl nitrite.

Circulating nitrite has recently emerged as an important physiological metabolite that contributes to increase vasodilation during tissue hypoxia. Using a wire myograph, we have investigated how the nitrite-dependent vasodilation in rat aortic rings is controlled by oxygen tension, norepinephrine...... are sufficient to induce NO-mediated vasodilation independently of the nitritereductase activities here investigated. These results further indicate that the vasoactive effect of nitrite is intrinsic to the vessel and may be due to S-nitrosothiols formed within the arterial smooth muscle....

Nitrite is widely consumed from the diet by animals and humans. However the largest contribution to exposure results from the in vivo conversion of exogenously derived nitrate to nitrite. Because of its potential to cause to methaemoglobin (MetHb) formation at excessive levels of intake, nitrite is regulated in feed and water as an undesirable substance. Forages and contaminated water have been shown to contain high levels of nitrate and represent the largest contributor to nitrite exposure for food-producing animals. Interspecies differences in sensitivity to nitrite intoxication principally result from physiological and anatomical differences in nitrite handling. In the case of livestock both pigs and cattle are relatively susceptible. With pigs this is due to a combination of low levels of bacterial nitritereductase and hence potential to reduce nitrite to ammonia as well as reduced capacity to detoxify MetHb back to haemoglobin (Hb) due to intrinsically low levels of MetHb reductase. In cattle the sensitivity is due to the potential for high dietary intake and high levels of rumen conversion of nitrate to nitrite, and an adaptable gut flora which at normal loadings shunts nitrite to ammonia for biosynthesis. However when this escape mechanism gets overloaded, nitrite builds up and can enter the blood stream resulting in methemoglobinemia. Looking at livestock case histories reported in the literature no-observed-effect levels of 3.3 mg/kg body weight (b.w.) per day for nitrite in pigs and cattle were estimated and related to the total daily nitrite intake that would result from complete feed at the EU maximum permissible level. This resulted in margins of safety of 9-fold and 5-fold for pigs and cattle, respectively. Recognising that the bulkiness of animal feed limits their consumption, these margins in conjunction with good agricultural practise were considered satisfactory for the protection of livestock health. A human health risk assessment was also

Full Text Available Cu(I-catalyzed azide-alkyne cycloaddition (CuAAC is an efficient reaction linking an azido and an alkynyl group in the presence of copper catalyst. Incorporation of a non-natural amino acid (NAA containing either an azido or an alkynyl group into a protein allows site-specific bioconjugation in mild conditions via CuAAC. Despite its great potential, bioconjugation of an enzyme has been hampered by several issues including low yield, poor solubility of a ligand, and protein structural/functional perturbation by CuAAC components. In the present study, we incorporated an alkyne-bearing NAA into an enzyme, murine dihydrofolate reductase (mDHFR, in high cell density cultivation of Escherichia coli, and performed CuAAC conjugation with fluorescent azide dyes to evaluate enzyme compatibility of various CuAAC conditions comprising combination of commercially available Cu(I-chelating ligands and reductants. The condensed culture improves the protein yield 19-fold based on the same amount of non-natural amino acid, and the enzyme incubation under the optimized reaction condition did not lead to any activity loss but allowed a fast and high-yield bioconjugation. Using the established conditions, a biotin-azide spacer was efficiently conjugated to mDHFR with retained activity leading to the site-specific immobilization of the biotin-conjugated mDHFR on a streptavidin-coated plate. These results demonstrate that the combination of reactive non-natural amino acid incorporation and the optimized CuAAC can be used to bioconjugate enzymes with retained enzymatic activity.

Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) is an efficient reaction linking an azido and an alkynyl group in the presence of copper catalyst. Incorporation of a non-natural amino acid (NAA) containing either an azido or an alkynyl group into a protein allows site-specific bioconjugation in mild conditions via CuAAC. Despite its great potential, bioconjugation of an enzyme has been hampered by several issues including low yield, poor solubility of a ligand, and protein structural/functional perturbation by CuAAC components. In the present study, we incorporated an alkyne-bearing NAA into an enzyme, murine dihydrofolate reductase (mDHFR), in high cell density cultivation of Escherichia coli, and performed CuAAC conjugation with fluorescent azide dyes to evaluate enzyme compatibility of various CuAAC conditions comprising combination of commercially available Cu(I)-chelating ligands and reductants. The condensed culture improves the protein yield 19-fold based on the same amount of non-natural amino acid, and the enzyme incubation under the optimized reaction condition did not lead to any activity loss but allowed a fast and high-yield bioconjugation. Using the established conditions, a biotin-azide spacer was efficiently conjugated to mDHFR with retained activity leading to the site-specific immobilization of the biotin-conjugated mDHFR on a streptavidin-coated plate. These results demonstrate that the combination of reactive non-natural amino acid incorporation and the optimized CuAAC can be used to bioconjugate enzymes with retained enzymatic activity.

Five new mononuclear mixed ligand complexes of the type [Cu(NCCH3)(dbdmp)](ClO4)2, [M(ONO)(dbdmp)]ClO4, [M(pz) (dbdmp)](ClO4)2 where M = Cu(II) and Co(II), pz = 3,5-dimethylpyrazole and dbdmp = N,N-diethyl-N‧,N‧-bis((3,5-dimethyl-1H-pyrazol-1-yl)methyl)ethane-1,2-diamine have been synthesized and characterized by physico-chemical and spectroscopy studies. The crystal structures of three copper(II) complexes [Cu(NCCH3)(dbdmp)](ClO4)2, [Cu(ONO)(dbdmp)]ClO4 and [Cu(pz)(dbdmp)](ClO4)2 have been determined by single crystal X-ray diffraction studies. Structural analyses reveal the geometry of [Cu(pz)(dbdmp)](ClO4)2 is distorted square pyramidal and other two copper(II) complexes have distorted trigonal bipyramidal geometry. Molecular composition of cobalt(II) complexes have been determined by mass spectral data. The EPR spectra of copper(II) complexes in frozen acetonitrile solution exhibit axial spectra, characteristic of dx2-y2 ground state. Electrochemical studies of copper(II) complexes using glassy carbon as working electrode in acetonitrile solution show Cu(II)/Cu(I) couple with quasi reversible electron transfer versus Ag/Ag+ reference electrode. Antimicrobial activity of all the synthesized complexes were investigated against two Gram positive and two Gram negative bacterial strains.

The objectives of the present investigations are to see the effects of biomass power plant effluent on the carbohydrates, aminoacids, nitrite and nitrite enzyme activitiesand proline of Arachis hypogaea L. vat TCGS 320 under controlled pot culture methods. Plants were cultivated with 25, 50, 75 and 100% of the effluent and a control without the effluent. The treatment of the crop with 25% of the effluent has shown stimulatory effect on all the biochemical parameters studied. Carbohydrates, starch, aminoacids, protein, nitrate and nitritereductase enzymatic activities have increased in 10, 15, 20 DAS (days after sowing). In 25 and 30 DAS all biochemical parameters have decreased, due to environmental factors (rainfall, temperature, humidity, etc.). The present investigation clearly indicated that the biomass power plant effluent has stimulatory effect on all the biochemical contents at lower concentration, and at higher concentration they have deleterious effects.

Full Text Available Apart from ATP synthesis mitochondria have many other functions, one being nitritereductase activity. NO released from nitrite has been shown to protect the heart from ischemia/reperfusion injury in a cGMP-dependent manner. However, the exact impact of mitochondria on the release of NO from nitrite in cardiomyocytes is not completely understood. Besides mitochondria, a number of non-mitochondrial metalloproteins have been suggested to facilitate this process. The aim of this study was to investigate the impact of mitochondria on the bioactivation of nitrite in HL-1 cardiomyocytes.The levels of nitrosyl complexes of hemoglobin (NO-Hb and cGMP levels were measured by electron spin resonance spectroscopy and enzyme immunoassay. In addition the formation of free NO was determined by confocal microscopy as well as intracellular nitrite and S-nitrosothiols by chemoluminescence analysis. NO was released from nitrite in cell culture in an oxygen dependent manner. Application of specific inhibitors of the respiratory chain, p450, NO synthases and xanthine oxidoreductase showed that all four enzymatic systems are involved in the release of NO, but more than 50% of NO is released via the mitochondrial pathway. Only NO released by mitochondria activated cGMP synthesis. Cardiomyocytes co-cultured with red blood cells (RBC competed with RBC for nitrite, but free NO was detected only in HL-1 cells suggesting that RBC are not a source of NO in this model. Apart from activation of cGMP synthesis, NO formed in HL-1 cells diffused out of the cells and formed NO-Hb complexes. In addition nitrite was converted by HL-1 cells to S-nitrosyl complexes. In HL-1 cardiomyocytes, several enzymatic systems are involved in nitrite reduction to NO but only the mitochondrial pathway of NO release activates cGMP synthesis. Our data suggest that this pathway may be a key regulator of myocardial contractility especially under hypoxic conditions.

In roots, nitrate assimilation is dependent upon a supply of reductant that is initially generated by oxidative metabolism including the pentose phosphate pathway (OPPP). The uptake of nitrite into the plastids and its subsequent reduction by nitritereductase (NiR) and glutamate synthase (GOGAT) ar

The systemic response to decreasing oxygen levels is hypoxic vasodilation. While this mechanism has been known for more than a century, the underlying cellular events have remained incompletely understood. Nitrite signaling is critically involved in vessel relaxation under hypoxia. This can be attributed to the presence of myoglobin in the vessel wall together with other potential nitritereductases, which generate nitric oxide, one of the most potent vasodilatory signaling molecules. Questions remain relating to the precise concentration of nitrite and the exact dose-response relations between nitrite and myoglobin under hypoxia. It is furthermore unclear whether regulatory mechanisms exist which balance this interaction. Nitrite tissue levels were similar across all species investigated. We then investigated the exact fractional myoglobin desaturation in an ex vivo approach when gassing with 1% oxygen. Within a short time frame myoglobin desaturated to 58±12%. Given that myoglobin significantly contributes to nitrite reduction under hypoxia, dose-response experiments using physiological to pharmacological nitrite concentrations were conducted. Along all concentrations, abrogation of myoglobin in mice impaired vasodilation. As reactive oxygen species may counteract the vasodilatory response, we used superoxide dismutase and its mimic tempol as well as catalase and ebselen to reduce the levels of reactive oxygen species during hypoxic vasodilation. Incubation of tempol in conjunction with catalase alone and catalase/ebselen increased the vasodilatory response to nitrite. Our study shows that modest hypoxia leads to a significant nitrite-dependent vessel relaxation. This requires the presence of vascular myoglobin for both physiological and pharmacological nitrite levels. Reactive oxygen species, in turn, modulate this vasodilation response.

The yield of nitric oxide from 1 mM sodium nitrite differs 200 times when the process was initiated by 10 mM sodium dithionite in the solution of 5 or 150 mM HEPES-buffer (pH 7.4). Dithionite acted both as a strong reductant and an agent that induced a local acidification of solutions without notable change in pH value. The amount of nitric oxide was estimated by the EPR method by measuring the incorporation of nitric oxide to water-soluble complexes of Fe with N-methyl-D-glucamine dithiocarbamate (MGD), which led to the formation of EPR-detectable mononitrosyl iron complexes with MGD (MNIC-MGD). Ten seconds after dithionite addition, the concentration of MNIC - MGD complexes reached 2 microM in 5 mM HEPES-buffer in contrast to 0.01 microM in 150 mM HEPES-buffer. The difference was suggested to be due to a higher life-time of zones with decreased pH values in a weaker weak buffer solution. The life-time was high enough to ensure the protonation of a part of nitrite. The resulting nitrous acid was decomposed to form nitric oxide. The difference in the formation of nitric oxide from nitrite was also observed in weak and strong buffer solutions in the presence of hemoglobin (0.3 mM) or serum albumin (0.5 mM). However, the ratios of nitric oxide yields in weak and strong buffer did not exceed 3-4 times. The increase in the formation of nitric oxide from nitrite was characteristic for the solutions containing both proteins. Large amounts of nitric oxide formed from nitrite was observed in mouse liver preparation subjected to freezing-thawing procedure followed by incubation in 150 mM HEPES-buffer (pH 7.4) and addition of dithionite. The proposition was made that the presence of zones with low pH value in cells and tissues can ensure the predominant operation of the acid mechanism formation of nitric oxide from nitrite. The contribution of the formation of nitric oxide from nitrite catalyzing with heme-containing proteins nitritereductases can be minor one under these

and nitrite on the O2 consumption rate and isometric twitch force development in electrically paced ventricular preparations during hypoxia, and measured O2 affinity and nitritereductase activity of the purified heart Mbs of both species. Upon hypoxia (9% O2), O2 consumption and developed force decreased...

The airway epithelium provides important barrier and host defense functions. Recent studies reveal that nitrite is an endocrine reservoir of nitric oxide (NO) bioactivity that is converted to NO by enzymatic reductases along the physiological oxygen gradient. Nitrite signaling has been described as NO dependent activation mediated by reactions with deoxygenated redox active hemoproteins, such as hemoglobin, myoglobin, neuroglobin, xanthine oxidoreductase (XO) and NO synthase at low pH and oxygen tension. However, nitrite can also be readily oxidized to nitrogen dioxide (NO(2)·) via heme peroxidase reactions, suggesting the existence of alternative oxidative signaling pathways for nitrite under normoxic conditions. In the present study, we examined normoxic signaling effects of sodium nitrite on airway epithelial cell wound healing. In an in vitro scratch injury model under normoxia, we exposed cultured monolayers of human airway epithelial cells to various concentrations of sodium nitrite and compared responses to NO donor. We found sodium nitrite potently enhanced airway epithelium wound healing at physiological concentrations (from 1 μM). The effect of nitrite was blocked by the NO and NO(2)· scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO). Interestingly, nitrite treatment did not increase cyclic guanosine monophosphate (cGMP) levels under these normoxic conditions, even in the presence of a phosphodiesterase 5 inhibitor, suggesting cGMP independent signaling. Consistent with an oxidative signaling pathway requiring hydrogen peroxide (H(2)O(2))/heme-peroxidase/NO(2)· signaling, the effects of nitrite were potentiated by superoxide dismutase (SOD) and low concentration H(2)O(2), whereas inhibited completely by catalase, followed by downstream extracellular-signal-regulated kinase (ERK) 1/2 activation. Our data represent the first description of normoxic nitrite signaling on lung epithelial cell proliferation and wound

Full Text Available Abstract Background Proteins having similar functions from different sources can be identified by the occurrence in their sequences, a conserved cluster of amino acids referred to as pattern, motif, signature or fingerprint. The wide usage of protein sequence analysis in par with the growth of databases signifies the importance of using patterns or signatures to retrieve out related sequences. Blue copper proteins are found in the electron transport chain of prokaryotes and eukaryotes. The signatures already existing in the databases like the type 1 copper blue, multiple copper oxidase, cyt b/b6, photosystem 1 psaA&B, psaG&K, and reiske iron sulphur protein are not specified signatures for blue copper proteins as the name itself suggests. Most profile and motif databases strive to classify protein sequences into a broad spectrum of protein families. This work describes the signatures designed based on the copper metal binding motifs in blue copper proteins. The common feature in all blue copper proteins is a trigonal planar arrangement of two nitrogen ligands [each from histidine] and one sulphur containing thiolate ligand [from cysteine], with strong interactions between the copper center and these ligands. Results Sequences that share such conserved motifs are crucial to the structure or function of the protein and this could provide a signature of family membership. The blue copper proteins chosen for the study were plantacyanin, plastocyanin, cucumber basic protein, stellacyanin, dicyanin, umecyanin, uclacyanin, cusacyanin, rusticyanin, sulfocyanin, halocyanin, azurin, pseudoazurin, amicyanin and nitritereductase which were identified in both eukaryotes and prokaryotes. ClustalW analysis of the protein sequences of each of the blue copper proteins was the basis for designing protein signatures or peptides. The protein signatures and peptides identified in this study were designed involving the active site region involving the amino acids

In the present study, we have constructed a bioluminescent bioreporter for the assessment of nitrate/nitrite bioavailability in wastewater. Specifically, an approximately 500-bp DNA fragment containing a nitrate/nitrite-activated nasR-like promoter (regulating expression of genes encoding nitritereductase in the genus Klebsiella) was fused upstream of the Vibrio fischeri luxCDABE gene cassette in a modified mini-Tn5 vector. Characterization of this strain, designated W6-1, yielded dose-dependent increased bioluminescence coincident with increased nitrate, nitrite, and ammonium added to the growth medium from 1 to 11 ppm. Bioluminescence in response to nitrogen species addition was light dependent up to 10, 7, and 8 ppm with nitrate, nitrite, and ammonium, respectively. This response was linear in the range from 1 to 8 ppm for nitrate (R2 = 0.98), 1 to 6 ppm for nitrite (R2 = 0.99), and 1 to 7 ppm for ammonium (R2 = 0.99). A significant bioluminescent response was also recorded when strain W6-1 was incubated with slurries from aged, nitrate/nitrite contaminated wastewater. Thus, bioreporter strain W6-1 can be used to elucidate factors that constrain the use of nitrate/nitrite in wastewaters.

Our long-time niche in synthetic biological inorganic chemistry has been to design ligands and generate coordination complexes of copper or iron ions or both, those reacting with dioxygen (O2) or nitrogen oxides (e.g., nitric oxide (NO(g)) and nitrite (NO2(-))) or both. As inspiration for this work, we turn to mitochondrial cytochrome c oxidase, which is responsible for dioxygen consumption and is also the predominant target for NO(g) and nitrite within mitochondria. In this Account, we highlight recent advances in studying synthetic heme/Cu complexes in two respects. First, there is the design, synthesis, and characterization of new O2 adducts whose further study will add insights into O2 reductive cleavage chemistry. Second, we describe how related heme/Cu constructs reduce nitrite ion to NO(g) or the reverse, oxidize NO(g) to nitrite. The reactions of nitrogen oxides occur as part of CcO's function, which is intimately tied to cellular O2 balance. We had first discovered that reduced heme/Cu compounds react with O2 giving μ-oxo heme-Fe(III)-O-Cu(II)(L) products; their properties are discussed. The O-atom is derived from dioxygen, and interrogations of these systems led to the construction and characterization of three distinctive classes of heme-peroxo complexes, two high-spin and one low-spin species. Recent investigations include a new approach to the synthesis of low-spin heme-peroxo-Cu complexes, employing a "naked" synthon, where the copper ligand denticity and geometric types can be varied. The result is a collection of such complexes; spectroscopic and structural features (by DFT calculations) are described. Some of these compounds are reactive toward reductants/protons effecting subsequent O-O cleavage. This points to how subtle improvements in ligand environment lead to a desired local structure and resulting optimized reactivity, as known to occur at enzyme active sites. The other sector of research is focused on heme/Cu assemblies mediating the redox

Full Text Available The main reason for high concentration of nitrite ions in water is the existence of sources of industrial and agricultural pollution. Contamination of drinking water, juices, wine and other liquids of nitrite ions as a result of improper use of nitrogen fertilizers has an adverse effect on living organism, because under the influence of enzymes nitrite ions in living organisms form high carcinogenic nitrosamines, and the interaction of nitrite ions from blood hemoglobin causes such toxicity that leads to disease cyanosis [1]. Therefore removal of nitrite ions from water has received increased attention. The paper discusses an innovative wastewater treatment technology from the nitrite ion with hypochlorite produced during electrolysis.

The main reason for high concentration of nitrite ions in water is the existence of sources of industrial and agricultural pollution. Contamination of drinking water, juices, wine and other liquids of nitrite ions as a result of improper use of nitrogen fertilizers has an adverse effect on living organism, because under the influence of enzymes nitrite ions in living organisms form high carcinogenic nitrosamines, and the interaction of nitrite ions from blood hemoglobin causes such toxicity t...

The nitritereductase activity of deoxyhemoglobin has received much recent interest because the nitric oxide produced in this reaction may participate in blood flow regulation during hypoxia. The present study used spectral deconvolution to characterize the reaction of nitrite with carp and rabbit...... hemoglobin at different constant oxygen tensions that generate the full range of physiological relevant oxygen saturations. Carp is a hypoxia-tolerant species with very high hemoglobin oxygen affinity, and the high R-state character and low redox potential of the hemoglobin is hypothesized to promote...... NO generation from nitrite. The reaction of nitrite with deoxyhemoglobin leads to a 1 : 1 formation of nitrosylhemoglobin and methemoglobin in both species. At intermediate oxygen saturations, the reaction with deoxyhemoglobin is clearly favored over that with oxyhemoglobin, and the oxyhemoglobin reaction...

Nitrite oxidation is an essential step in transformations of fixed nitrogen. The physiology of nitrite oxidizing bacteria (NOB) implies that the rates of nitrite oxidation should be controlled by concentration of their substrate, nitrite, and the terminal electron acceptor, oxygen. The sensitivities of nitrite oxidation to oxygen and nitrite concentrations were investigated using 15N tracer incubations in the Eastern Tropical North Pacific. Nitrite stimulated nitrite oxidation under low in situ nitrite conditions, following Michaelis-Menten kinetics, indicating that nitrite was the limiting substrate. The nitrite half-saturation constant (Ks = 0.254 ± 0.161 μM) was 1-3 orders of magnitude lower than in cultivated NOB, indicating higher affinity of marine NOB for nitrite. The highest rates of nitrite oxidation were measured in the oxygen depleted zone (ODZ), and were partially inhibited by additions of oxygen. This oxygen sensitivity suggests that ODZ specialist NOB, adapted to low-oxygen conditions, are responsible for apparently anaerobic nitrite oxidation.

Nitrite is an important species in the global nitrogen cycle, and the nitritereductase enzymes convert nitrite to nitric oxide (NO). Recently, it has been shown that hemoglobin and myoglobin catalyze the reduction of nitrite to NO under hypoxic conditions. We have determined the 1.20 Angstroms resolution crystal structure of the nitrite adduct of ferric horse heart myoglobin (hh Mb). The ligand is bound to iron in the nitrito form, and the complex is formulated as Mb{sup III}(ONO{sup -}). The Fe-ONO bond length is 1.94 Angstroms, and the O-N-O angle is 113 degrees. In addition, the nitrite ligand is stabilized by hydrogen bonding with the distal His64 residue. We have also determined the 1.30 Angstroms resolution crystal structures of hh Mb{sup II}NO. When hh Mb{sup II}NO is prepared from the reaction of metMb{sup III} with nitrite/dithionite, the FeNO angle is 144 degrees with a Fe-NO bond length of 1.87 Angstroms. However, when prepared from the reaction of NO with reduced Mb{sup II}, the FeNO angle is 120 degrees with a Fe-NO bond length of 2.13 Angstroms. This difference in FeNO conformations as a function of preparative method is reproducible, and suggests a role of the distal pocket in hh Mb{sup II}NO in stabilizing local FeNO conformational minima.

Full Text Available Nitric oxide (NO is a key regulator of vascular tone. Endothelial nitric oxide synthase (eNOS is responsible for NO generation under normoxic conditions. Under hypoxia however, eNOS is inactive and red blood cells (RBC provide an alternative NO generation pathway from nitrite to regulate hypoxic vasodilation. While nitritereductase activity of hemoglobin is well acknowledged, little is known about generation of NO by intact RBC with physiological hemoglobin concentrations. We aimed to develop and apply a new approach to provide insights in the ability of RBC to convert nitrite into NO under hypoxic conditions. We established a novel experimental setup to evaluate nitrite uptake and the release of NO from RBC into the gas-phase under different conditions. NO measurements were similar to well-established clinical measurements of exhaled NO. Nitrite uptake was rapid, and after an initial lag phase NO release from RBC was constant in time under hypoxic conditions. The presence of oxygen greatly reduced NO release, whereas inhibition of eNOS and xanthine oxidoreductase (XOR did not affect NO release. A decreased pH increased NO release under hypoxic conditions. Hypothermia lowered NO release, while hyperthermia increased NO release. Whereas fetal hemoglobin did not alter NO release compared to adult hemoglobin, sickle RBC showed an increased ability to release NO. Under all conditions nitrite uptake by RBC was similar. This study shows that nitrite uptake into RBC is rapid and release of NO into the gas-phase continues for prolonged periods of time under hypoxic conditions. Changes in the RBC environment such as pH, temperature or hemoglobin type, affect NO release.

Nitric oxide (NO) is a key regulator of vascular tone. Endothelial nitric oxide synthase (eNOS) is responsible for NO generation under normoxic conditions. Under hypoxia however, eNOS is inactive and red blood cells (RBC) provide an alternative NO generation pathway from nitrite to regulate hypoxic vasodilation. While nitritereductase activity of hemoglobin is well acknowledged, little is known about generation of NO by intact RBC with physiological hemoglobin concentrations. We aimed to develop and apply a new approach to provide insights in the ability of RBC to convert nitrite into NO under hypoxic conditions. We established a novel experimental setup to evaluate nitrite uptake and the release of NO from RBC into the gas-phase under different conditions. NO measurements were similar to well-established clinical measurements of exhaled NO. Nitrite uptake was rapid, and after an initial lag phase NO release from RBC was constant in time under hypoxic conditions. The presence of oxygen greatly reduced NO release, whereas inhibition of eNOS and xanthine oxidoreductase (XOR) did not affect NO release. A decreased pH increased NO release under hypoxic conditions. Hypothermia lowered NO release, while hyperthermia increased NO release. Whereas fetal hemoglobin did not alter NO release compared to adult hemoglobin, sickle RBC showed an increased ability to release NO. Under all conditions nitrite uptake by RBC was similar. This study shows that nitrite uptake into RBC is rapid and release of NO into the gas-phase continues for prolonged periods of time under hypoxic conditions. Changes in the RBC environment such as pH, temperature or hemoglobin type, affect NO release.

Urinary tract infections (UTIs) are the most common type of nosocomial infection. Traditionally, the presence of white blood cells and microorganisms in the urine provides objective evidence for UTI diagnosis. Here, we describe the use of liquid chromatography-tandem mass spectrometry (LC-MS/MS) to measure the nitrite and nitrate levels in urine and investigate the potential of this method for UTI diagnosis. LC-MS/MS analysis was performed in positive electrospray ionization mode. After adding (15)N-labeled internal standards and derivatizing with 2,3-diaminonaphthalene (DAN), the urinary nitrite content was directly analyzed by LC-MS/MS, whereas the urinary nitrate was first reduced to nitrite before derivatization and LC-MS/MS analysis. The derivatization of nitrite and enzymatic reduction of nitrate were optimized. This method was then applied to 241 healthy subjects and 73 UTI patients. Optimization tests revealed that 1 mL of crude urine required at least 6.25 μmol of DAN to completely derivatize nitrite and 2.5 U of nitrate reductase to completely reduce nitrate to nitrite. Urinary analysis showed that the urinary concentration of nitrite and the nitrite/nitrate ratio were higher in UTI patients than in healthy subjects. Compared with the dipstick-based urinary nitrite test and using LC-MS/MS to determine the nitrite concentration (sensitivity: 23-25%), the nitrite/nitrate ratio was significantly more sensitive (95%) and exhibited a satisfactory specificity (91%) in the screening of UTIs. Taken together, the nitrite/nitrate ratio, which reflects the reducing ability of pathogenic bacteria, could be a better method for the diagnosis of UTIs that is not subject to variations in urine specimen quality.

The periplasmic nitritereductase system from Escherichia coli and the extracellular Fe(III) reductase system from Shewanella oneidensis contain multihaem c-type cytochromes as electron carriers and terminal reductases. The position and orientation of the haem cofactors in multihaem cytochromes from different bacteria often show significant conservation despite different arrangements of the polypeptide chain. We propose that the decahaem cytochromes of the iron reductase system MtrA, MtrC and OmcA comprise pentahaem 'modules' similar to the electron donor protein, NrfB, from E. coli. To demonstrate this, we have isolated and characterized the N-terminal pentahaem module of MtrA by preparing a truncated form containing five covalently attached haems. UV-visible spectroscopy indicated that all five haems were low-spin, consistent with the presence of bis-His ligand co-ordination as found in full-length MtrA.

The functions of quinone reductase 2 have eluded researchers for decades even though a genetic polymorphism is associated with various neurological disorders. Employing enzymatic studies using adrenochrome as a substrate, we show that quinone reductase 2 is specific for the reduction of adrenochrome, whereas quinone reductase 1 shows no activity. We also solved the crystal structure of quinone reductase 2 in complexes with dopamine and adrenochrome, two compounds that are structurally related to catecholamine quinones. Detailed structural analyses delineate the mechanism of quinone reductase 2 specificity toward catechol quinones in comparison with quinone reductase 1; a side-chain rotational difference between quinone reductase 1 and quinone reductase 2 of a single residue, phenylalanine 106, determines the specificity of enzymatic activities. These results infer functional differences between two homologous enzymes and indicate that quinone reductase 2 could play important roles in the regulation of catecholamine oxidation processes that may be involved in the etiology of Parkinson disease.

Full Text Available The health benefits of nitrite are now indisputable when administered in a clinical setting for specific diseases. Currently, most published reports identify the production of nitric oxide (NO as the mechanism of action for nitrite. Basic science, in addition to clinical studies, demonstrate that nitrite and/or nitrate cannot restore NO homeostasis as an endothelium independent source of NO that may be a redundant system for endogenous NO production. Nitrate must first be reduced to nitrite by oral commensal bacteria; nitrite can then be further reduced to NO along the physiological oxygen gradient. But despite decades of rigorous research on sodium nitrate’s safety and efficacy as a curing agent, sodium nitrite is still regarded by many as a toxic undesirable food additive. However, research within the biomedical science community has revealed enormous therapeutic benefits of nitrite which are being developed as novel therapies for conditions associated with nitric oxide insufficiency. Thus, this review will highlight the fundamental biochemistry of nitrite in human physiology and provide evidence that nitrite be considered an essential nutrient. Foods or diets enriched with nitrite can have profound positive health benefits.

The proposal that nitrite exerts its inhibitory effect on anaerobic bacteria by direct interaction with the iron-sulphur proteins of the phosphoroclastic system was investigated. The effects of nitrate, nitrite with or without ascorbate, and nitric oxide on the growth of Clostridium sporogenes in liquid cultures at pH 7.4, on the rates of hydrogen production, and on the activities of the enzymes pyruvate-ferredoxin oxidoreductase and hydrogenase, and of ferredoxin were investigated. In agreement with previous studies, nitrate was the least effective inhibitor of cell growth, and nitric oxide the most effective. Nitritereductase activity was very low in C. sporogenes, indicating that the presence of external reducing agents would be necessary for the reduction of nitrite to nitric oxide. Inhibition by nitrite was enhanced by ascorbate; 0.5 mM-nitrite with 10 mM-ascorbate stopped growth completely. In partially-purified preparations 4.1 mM-NaNO2 and equimolar ascorbate caused complete inactivation of hydrogenase activity but only partial (up to 78%) inactivation of pyruvate-ferredoxin oxidoreductase. This agreed with the loss of hydrogen production observed with nitrite in vivo. Inhibition occurred within 5 min, and was irreversible in each case. Electron paramagnetic resonance (EPR) spectroscopy showed that paramagnetic [Fe(NO)2(SR)2] species were formed during growth in the presence of nitrite, and were associated with cells. However, the intensity of these EPR signals did not correlate with the inhibition of cell growth. The [4Fe-4S] clusters in ferredoxin were shown by EPR spectroscopy to be resistant to treatment with 3.6 mM-NaNO2 and 3.6 mM-ascorbate. It is concluded that the effects of nitrite on pre-formed iron-sulphur proteins are not convincing as a basis for the lethal effects on bacterial cells.

In order to select the strain that can degrade nitrite,we use the screening plate with nitrite as the sole nitrogen source to select the strain with ability to degrade nitrite,and get a strain with nitrite degrading capacity from the silt of shrimp farming pond in Hepu City,Guangxi Zhuang Autonomous Region. By identifying the strain from colony morphology,physiological and biochemical characteristics and 16 S r RNA sequence,we finally get a bacteria strain that can degrade nitrite,and this strain can grow well on the culture medium with nitrite concentration of 2 g / L. Based on morphology,nitrogen source requirements and evolutionary tree analysis of the above 16 S r RNA sequence,it is found that this strain belongs to Pannonibacter phragmitetus. According to the screening location,it is named HPPP007 strain.

Vascular relaxation to nitroglycerin (glyceryl trinitrate; GTN) requires its bioactivation by mechanisms that remain controversial. We report here that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the release of nitrite from GTN. In assays containing dithiothreitol (DTT) and NAD(+), the GTN reductase activity of purified GAPDH produces nitrite and 1,2-GDN as the major products. A vmax of 2.6nmolmin(-)(1)mg(-)(1) was measured for nitrite production by GAPDH from rabbit muscle and a GTN KM of 1.2mM. Reductive denitration of GTN in the absence of DTT results in dose- and time-dependent inhibition of GAPDH dehydrogenase activity. Disulfiram, a thiol-modifying drug, inhibits both the dehydrogenase and GTN reductase activity of GAPDH, while DTT or tris(2-carboxyethyl)phosphine reverse the GTN-induced inhibition. Incubation of intact human erythrocytes or hemolysates with 2mM GTN for 60min results in 50% inhibition of GAPDH's dehydrogenase activity, indicating that GTN is taken up by these cells and that the dehydrogenase is a target of GTN. Thus, erythrocyte GAPDH may contribute to GTN bioactivation.

Full Text Available Nitrate reductase catalyses the oxidation of NAD(PH and the reduction of nitrate to nitrite. NR serves as a central point for the integration of metabolic pathways by governing the flux of reduced nitrogen through several regulatory mechanisms in plants, algae and fungi. Bacteria express nitrate reductases that convert nitrate to nitrite, but mammals lack these specific enzymes. The microbial nitrate reductase reduces toxic compounds to nontoxic compounds with the help of NAD(PH. In the present study, our results revealed that Bacillus weihenstephanensis expresses a nitrate reductase enzyme, which was made to generate the 3D structure of the enzyme. Six different modelling servers, namely Phyre2, RaptorX, M4T Server, HHpred, SWISS MODEL and Mod Web, were used for comparative modelling of the structure. The model was validated with standard parameters (PROCHECK and Verify 3D. This study will be useful in the functional characterization of the nitrate reductase enzyme and its docking with nitrate molecules, as well as for use with autodocking.

... Listing § 573.700 Sodium nitrite. Sodium nitrite may be safely used in canned pet food containing meat and fish in accordance with the following prescribed conditions: (a) It is used or intended for use alone as a preservative and color fixative in canned pet food containing fish, meat, and fish and...

Copper(II) is known to catalyze the generation of reactive nitrogen species in the presence of hydrogen peroxide, nitrite or nitric oxide, leading to tyrosine nitration, a biomarker for free radical species associated diseases. Here, we find that biological antioxidants such as ascorbic acid can promote tyrosine nitration in the presence of copper(II) and nitrite under aerobic and weak acidic conditions. Tyrosine nitration is demonstrated on both β-amyloid peptide and angiotensin I. These stu...

The alphaproteobacterium Nitrobacter winogradskyi (ATCC 25391) is a gram-negative facultative chemolithoautotroph capable of extracting energy from the oxidation of nitrite to nitrate. Sequencing and analysis of its genome revealed a single circular chromosome of 3,402,093 bp encoding 3,143 predicted proteins. There were extensive similarities to genes in two alphaproteobacteria, Bradyrhizobium japonicum USDA110 (1,300 genes) and Rhodopseudomonas palustris CGA009 CG (815 genes). Genes encoding pathways for known modes of chemolithotrophic and chemoorganotrophic growth were identified. Genes encoding multiple enzymes involved in anapleurotic reactions centered on C2 to C4 metabolism, including a glyoxylate bypass, were annotated. The inability of N. winogradskyi to grow on C6 molecules is consistent with the genome sequence, which lacks genes for complete Embden-Meyerhof and Entner-Doudoroff pathways, and active uptake of sugars. Two gene copies of the nitrite oxidoreductase, type I ribulose-1,5-bisphosphate carboxylase/oxygenase, cytochrome c oxidase, and gene homologs encoding an aerobic-type carbon monoxide dehydrogenase were present. Similarity of nitrite oxidoreductases to respiratory nitrate reductases was confirmed. Approximately 10% of the N. winogradskyi genome codes for genes involved in transport and secretion, including the presence of transporters for various organic-nitrogen molecules. The N. winogradskyi genome provides new insight into the phylogenetic identity and physiological capabilities of nitrite-oxidizing bacteria. The genome will serve as a model to study the cellular and molecular processes that control nitrite oxidation and its interaction with other nitrogen-cycling processes.

The first examples of inorganic nitrite complexes of the natural actinides are described, including the structures of the homoleptic thorium(IV) [PPh(4)](2)[Th(NO(2))(6)] and the uranyl(VI) [PPh(4)](2)[UO(2)(NO(2))(4)] complexes; the nitrite ligand can adopt two different coordination modes in the coordination sphere of the uranyl ion and is unstable towards reduction.

The variables of direct importance in the reduction of nitrate to nitrite by a metallic reductant such as cadmium used in a reductor column are discussed with special reference to the determination of nitrate as nitrite in very dilute solutions, e.g., natural waters. As a result of these considerations the effect of flow-rate (expressed as bed-volumes min ), pH, temperature, chloride concentration and various types of reductor cadmium on the yield of nitrite is investigated. The effect of dissolved oxygen in the sample solution on pH and cadmium concentration in the reduced solution is demonstrated. At constant pH a maximum yield of nitrite is obtained at a certain flow-rate, which is explained as the result of a rapid formation and simultaneously proceeding slow reduction of nitrite. With increasing pH this maximum is shifted to lower flow-rates, and grows broader whilst the yield at maximum approaches 100%; at pH 9.5 a yield of 99.9 +/- 0.1% is obtained. The temperature has little effect on the reduction rate in the interval 20-30 degrees but at 10 degrees the reduction is noticeably slower. Chloride ions have a strongly retarding effect on the reduction rate but the yield at maximum is not affected. Electrolytically precipitated cadmium, filings of pure cadmium or amalgamated pure cadmium all give practically the same yield at maximum though some differences in reduction rate are observed. Impure cadmium or copper-cadmium and silver-cadmium, owing to the formation of galvanic cells with higher reducing power, give a high reduction rate, which also applies to nitrite, causing a poorer yield at maximum. The practical consequences of the results are thoroughly discussed.

A specific form of plasma membrane-bound nitrate reductase in plants is restricted to roots. Two peptides originated from plasma membrane integral proteins isolated from Hordeum vulgare have been assigned as homologues to the subunit NarH of respiratory nitrate reductase of Escherichia coli. Corresponding sequences have been detected for predicted proteins of Populus trichocarpa with high degree of identities for the subunits NarH (75%) and NarG (65%), however, with less accordance for the subunit NarI. These findings coincide with biochemical properties, particularly in regard to the electron donors menadione and succinate. Together with the root-specific and plasma membrane-bound nitrite/NO reductase, nitric oxide is produced under hypoxic conditions in the presence of nitrate. In this context, a possible function in nitrate respiration of plant roots and an involvement of plants in denitrification processes are discussed.

Nitric oxide (NO) is a signaling and defense molecule of major importance in living organisms. In the model legume Medicago truncatula, NO production has been detected in the nitrogen fixation zone of the nodule, but the systems responsible for its synthesis are yet unknown and its role in symbiosis is far from being elucidated. In this work, using pharmacological and genetic approaches, we explored the enzymatic source of NO production in M. truncatula-Sinorhizobium meliloti nodules under normoxic and hypoxic conditions. When transferred from normoxia to hypoxia, nodule NO production was rapidly increased, indicating that NO production capacity is present in functioning nodules and may be promptly up-regulated in response to decreased oxygen availability. Contrary to roots and leaves, nodule NO production was stimulated by nitrate and nitrite and inhibited by tungstate, a nitrate reductase inhibitor. Nodules obtained with either plant nitrate reductase RNA interference double knockdown (MtNR1/2) or bacterial nitrate reductase-deficient (napA) and nitritereductase-deficient (nirK) mutants, or both, exhibited reduced nitrate or nitritereductase activities and NO production levels. Moreover, NO production in nodules was found to be inhibited by electron transfer chain inhibitors, and nodule energy state (ATP-ADP ratio) was significantly reduced when nodules were incubated in the presence of tungstate. Our data indicate that both plant and bacterial nitrate reductase and electron transfer chains are involved in NO synthesis. We propose the existence of a nitrate-NO respiration process in nodules that could play a role in the maintenance of the energy status required for nitrogen fixation under oxygen-limiting conditions.

Among vertebrates able to tolerate periods of oxygen deprivation, the painted and red-eared slider turtles (Chrysemys picta and Trachemys scripta) and the crucian carp (Carassius carassius) are the most extreme and can survive even months of total lack of oxygen during winter. The key to hypoxia ...... of NO and nitrite signaling in the adaptive response to hypoxia in vertebrate animals....... survival resides in concerted physiological responses, including strong metabolic depression, protection against oxidative damage and – in air breathing animals - redistribution of blood flow. Each of these responses is known to be tightly regulated by nitric oxide (NO) and during hypoxia by its metabolite...... nitrite. The aim of this review is to highlight recent work illustrating the widespread roles of NO and nitrite in the tolerance to extreme oxygen deprivation, in particular in the red-eared slider turtle and crucian carp, but also in diving marine mammals. The emerging picture underscores the importance...

In mammals, treatment with low doses of nitrite has a cytoprotective effect in ischemia/reperfusion events, as a result of nitric oxide formation and S-nitrosation of proteins. Interestingly, anoxia-tolerant lower vertebrates possess an intrinsic ability to increase intracellular nitrite concentr......In mammals, treatment with low doses of nitrite has a cytoprotective effect in ischemia/reperfusion events, as a result of nitric oxide formation and S-nitrosation of proteins. Interestingly, anoxia-tolerant lower vertebrates possess an intrinsic ability to increase intracellular nitrite...... concentration during anoxia in tissues with high myoglobin and mitochondria content, such as the heart. Here, we tested the hypothesis that red and white skeletal muscles develop different nitrite levels in crucian carp exposed to deep hypoxia and assessed whether this correlates with myoglobin concentration....... We also tested whether liver, muscle and heart tissue possess nitrate reductase activity that supplies nitrite to the tissues during severe hypoxia. Crucian carp exposed to deep hypoxia (1

Full Text Available BACKGROUND: Nitrite is a nitric oxide (NO metabolite in tissues and blood, which can be converted to NO under hypoxia to facilitate tissue perfusion. Although nitrite is known to cause vasodilation following its reduction to NO, the effect of nitrite on platelet activity remains unclear. In this study, the effect of nitrite and nitrite+erythrocytes, with and without deoxygenation, on platelet activity was investigated. METHODOLOGY/FINDING: Platelet aggregation was studied in platelet-rich plasma (PRP and PRP+erythrocytes by turbidimetric and impedance aggregometry, respectively. In PRP, DEANONOate inhibited platelet aggregation induced by ADP while nitrite had no effect on platelets. In PRP+erythrocytes, the inhibitory effect of DEANONOate on platelets decreased whereas nitrite at physiologic concentration (0.1 µM inhibited platelet aggregation and ATP release. The effect of nitrite+erythrocytes on platelets was abrogated by C-PTIO (a membrane-impermeable NO scavenger, suggesting an NO-mediated action. Furthermore, deoxygenation enhanced the effect of nitrite as observed from a decrease of P-selectin expression and increase of the cGMP levels in platelets. The ADP-induced platelet aggregation in whole blood showed inverse correlations with the nitrite levels in whole blood and erythrocytes. CONCLUSION: Nitrite alone at physiological levels has no effect on platelets in plasma. Nitrite in the presence of erythrocytes inhibits platelets through its reduction to NO, which is promoted by deoxygenation. Nitrite may have role in modulating platelet activity in the circulation, especially during hypoxia.

Shewanella putrefaciens W3-18-1 harbours two periplasmic nitrate reductase (Nap) gene clusters, NapC-associated nap-alpha (napEDABC) and CymA-dependent nap-beta (napDAGHB), for dissimilatory nitrate respiration. CymA is a member of the NapC/NirT quinol dehydrogenase family and acts as a hub to support different respiratory pathways, including those on iron [Fe(III)] and manganese [Mn(III, IV)] (hydr)oxide, nitrate, nitrite, fumarate and arsenate in Shewanella strains. However, in our analysis it was shown that another NapC/NirT family protein, NapC, was only involved in nitrate reduction, although both CymA and NapC can transfer quinol-derived electrons to a periplasmic terminal reductase or an electron acceptor. Furthermore, our results showed that NapC could only interact specifically with the Nap-alpha nitrate reductase while CymA could interact promiscuously with Nap-alpha, Nap-beta and the NrfA nitritereductase for nitrate and nitrite reduction. To further explore the difference in specificity, site-directed mutagenesis on both CymA and NapC was conducted and the phenotypic changes in nitrate and nitrite reduction were tested. Our analyses demonstrated that the Lys-91 residue played a key role in nitrate reduction for quinol oxidation and the Asp-166 residue might influence the maturation of CymA. The Asp-97 residue might be one of the key factors that influence the interaction of CymA with the cytochromes NapB and NrfA.

Shewanella putrefaciens W3-18-1 harbours two periplasmic nitrate reductase (Nap) gene clusters, NapC-associated nap-alpha (napEDABC) and CymA-dependent nap-beta (napDAGHB), for dissimilatory nitrate respiration. CymA is a member of the NapC/NirT quinol dehydrogenase family and acts as a hub to support different respiratory pathways, including those on iron [Fe(III)] and manganese [Mn(III, IV)] (hydr)oxide, nitrate, nitrite, fumarate and arsenate in Shewanella strains. However, in our analysis it was shown that another NapC/NirT family protein, NapC, was only involved in nitrate reduction, although both CymA and NapC can transfer quinol-derived electrons to a periplasmic terminal reductase or an electron acceptor. Furthermore, our results showed that NapC could only interact specifically with the Nap-alpha nitrate reductase while CymA could interact promiscuously with Nap-alpha, Nap-beta and the NrfA nitritereductase for nitrate and nitrite reduction. To further explore the difference in specificity, site-directed mutagenesis on both CymA and NapC was conducted and the phenotypic changes in nitrate and nitrite reduction were tested. Our analyses demonstrated that the Lys-91 residue played a key role in nitrate reduction for quinol oxidation and the Asp-166 residue might influence the maturation of CymA. The Asp-97 residue might be one of the key factors that influence the interaction of CymA with the cytochromes NapB and NrfA.

Polysiloxanes with different types of polar substituents are excellent membrane materials for nitrite and fluoride selective chemically modified field effect transistors (CHEMFETs). Nitrite selectivity has been introduced by incorporation of a cobalt porphyrin into the membrane; fluoride selectivity

Polysiloxanes with different types of polar substituents are excellent membrane materials for nitrite and fluoride selective chemically modified field effect transistors (CHEMFETs). Nitrite selectivity has been introduced by incorporation of a cobalt porphyrin into the membrane; fluoride selectivity

Bacteria that metabolize p-nitrophenol (PNP) oxidize the substrate to 3-ketoadipic acid via either hydroquinone or 1,2,4-trihydroxybenzene (THB); however, initial steps in the pathway for PNP biodegradation via THB are unclear. The product of initial hydroxylation of PNP could be either 4-nitrocatechol or 4-nitroresorcinol. Here we describe the complete pathway for aerobic PNP degradation by Bacillus sphaericus JS905 that was isolated by selective enrichment from an agricultural soil in India. Washed cells of PNP-grown JS905 released nitrite in stoichiometric amounts from PNP and 4-nitrocatechol. Experiments with extracts obtained from PNP-grown cells revealed that the initial reaction is a hydroxylation of PNP to yield 4-nitrocatechol. 4-Nitrocatechol is subsequently oxidized to THB with the concomitant removal of the nitro group as nitrite. The enzyme that catalyzed the two sequential monooxygenations of PNP was partially purified and separated into two components by anion-exchange chromatography and size exclusion chromatography. Both components were required for NADH-dependent oxidative release of nitrite from PNP or 4-nitrocatechol. One of the components was identified as a reductase based on its ability to catalyze the NAD(P)H-dependent reduction of 2,6-dichlorophenolindophenol and nitroblue tetrazolium. Nitrite release from either PNP or 4-nitrocatechol was inhibited by the flavoprotein inhibitor methimazole. Our results indicate that the two monooxygenations of PNP to THB are catalyzed by a single two-component enzyme system comprising a flavoprotein reductase and an oxygenase.

Bacteria that metabolize p-nitrophenol (PNP) oxidize the substrate to 3-ketoadipic acid via either hydroquinone or 1,2,4-trihydroxybenzene (THB); however, initial steps in the pathway for PNP biodegradation via THB are unclear. The product of initial hydroxylation of PNP could be either 4-nitrocatechol or 4-nitroresorcinol. Here the authors describe the complete pathway for aerobic PNP degradation by Bacillus sphaericus JS905 that was isolated by selective enrichment from an agricultural soil in India. Washed cells of PNP-grown JS905 released nitrite in stoichiometric amounts from PNP and 4-nitrocatechol. Experiments with extracts obtained from PNP-grown cells revealed that the initial reaction is a hydroxylation of PNP to yield 4-nitrocatechol. 4-nitrocatechol is subsequently oxidized to THB with the concomitant removal of the nitro group as nitrite. The enzyme that catalyzed the two sequential monooxygenations of PNP was partially purified and separated into two components by anion-exchange chromatography and size exclusion chromatography. Both components were required for NADH-dependent oxidative release of nitrite from PNP or 4-nitrocatechol. One of the components was identified as a reductase based on its ability to catalyze the NAD(P)H-dependent reduction of 2,6-dichlorophenolindophenol and nitroblue tetrazolium. Nitrite release from either PNP or 4-nitrocatechol was inhibited by the flavoprotein inhibitor methimazole. Their results indicate that the two monooxygenations of PNP to THB are catalyzed by a single two-component enzyme system comprising a flavoprotein reductase and an oxygenase.

nitrite-induced vasodilation (possibly via nitric oxide generated from nitrite) that is countered by increased cardiac pumping to re-establish blood pressure. Nitrite can form and/or mimic nitric oxide and thereby interfere with processes regulated by this local hormone. Steroid hormone synthesis may...... species and in some cases also within species. Rainbow trout fall into two groups with regard to susceptibility and physiological response. These two groups are not related to sex but show significant different nitrite uptake rates....

There are 2 nitrite maxima in the Northern Arabian Sea, one at the thermocline depth and the other at depths between 300 and 500 m. The 2nd maximum is more prominent in the northeastern part of the Arabian Sea. The 1st maximum is associated...

of the density and orientational distribution of NrfA molecules are disclosed. The submonolayer coverage resolved by in situ STM is readily reconciled with the failure to detect nonturnover signals in cyclic voltammetry of the NrfA films. The molecular structures show a range of lateral dimensions...... a direct approach to correlate electrocatalytic and molecular properties of the protein layer, a long-standing issue in protein film voltammetry....

, internal electron transfer between these sites is an inherent element in the catalytic cycle of this enzyme. We have investigated the internal electron transfer reaction employing pulse radiolytically produced N-methyl nicotinamide radicals as reductant which reacts solely with the heme-c in an essentially...... diffusion controlled process. Following this initial step, the reduction equivalent is equilibrating between the c and d(1) heme sites in a unimolecular process (k=23 s(-1), 298 K, pH 7.0) and an equilibrium constant of 1.0. The temperature dependence of this internal electron transfer process has been...

Nitrite oxidation is the second step of nitrification. It is the primary source of oceanic nitrate, the predominant form of bioavailable nitrogen in the ocean. Despite its obvious importance, nitrite oxidation has rarely been investigated in marine settings. We determined nitrite oxidation rates directly in (15)N-incubation experiments and compared the rates with those of nitrate reduction to nitrite, ammonia oxidation, anammox, denitrification, as well as dissimilatory nitrate/nitrite reduction to ammonium in the Namibian oxygen minimum zone (OMZ). Nitrite oxidation (≤372 nM NO(2)(-) d(-1)) was detected throughout the OMZ even when in situ oxygen concentrations were low to non-detectable. Nitrite oxidation rates often exceeded ammonia oxidation rates, whereas nitrate reduction served as an alternative and significant source of nitrite. Nitrite oxidation and anammox co-occurred in these oxygen-deficient waters, suggesting that nitrite-oxidizing bacteria (NOB) likely compete with anammox bacteria for nitrite when substrate availability became low. Among all of the known NOB genera targeted via catalyzed reporter deposition fluorescence in situ hybridization, only Nitrospina and Nitrococcus were detectable in the Namibian OMZ samples investigated. These NOB were abundant throughout the OMZ and contributed up to ~9% of total microbial community. Our combined results reveal that a considerable fraction of the recently recycled nitrogen or reduced NO(3)(-) was re-oxidized back to NO(3)(-) via nitrite oxidation, instead of being lost from the system through the anammox or denitrification pathways.

bicarbonate and nitrite, we hypothesized that CA uses nitrite as a substrate to produce the potent vasodilator nitric oxide (NO) to increase local blood flow to metabolically active tissues. Here we show that CA readily reacts with nitrite to generate NO, particularly at low pH, and that the NO produced...

The world production of copper is steadily increasing. Although humans are widely exposed to copper-containing items on the skin and mucosa, allergic reactions to copper are only infrequently reported. To review the chemistry, biology and accessible data to clarify the implications of copper hypersensitivity, a database search of PubMed was performed with the following terms: copper, dermatitis, allergic contact dermatitis, contact hypersensitivity, contact sensitization, contact allergy, patch test, dental, IUD, epidemiology, clinical, and experimental. Human exposure to copper is relatively common. As a metal, it possesses many of the same qualities as nickel, which is a known strong sensitizer. Cumulative data on subjects with presumed related symptoms and/or suspected exposure showed that a weighted average of 3.8% had a positive patch test reaction to copper. We conclude that copper is a very weak sensitizer as compared with other metal compounds. However, in a few and selected cases, copper can result in clinically relevant allergic reactions.

In adult humans, the net absorption of dietary copper is approximately 1 mg/d. Dietary copper joins some 4-5 mg of endogenous copper flowing into the gastrointestinal tract through various digestive juices. Most of this copper returns to the circulation and to the tissues (including liver) that formed them. Much lower amounts of copper flow into and out of other major parts of the body (including heart, skeletal muscle, and brain). Newly absorbed copper is transported to body tissues in two phases, borne primarily by plasma protein carriers (albumin, transcuprein, and ceruloplasmin). In the first phase, copper goes from the intestine to the liver and kidney; in the second phase, copper usually goes from the liver (and perhaps also the kidney) to other organs. Ceruloplasmin plays a role in this second phase. Alternatively, liver copper can also exit via the bile, and in a form that is less easily reabsorbed. Copper is also present in and transported by other body fluids, including those bathing the brain and central nervous system and surrounding the fetus in the amniotic sac. Ceruloplasmin is present in these fluids and may also be involved in copper transport there. The concentrations of copper and ceruloplasmin in milk vary with lactational stage. Parallel changes occur in ceruloplasmin messenger RNA expression in the mammary gland (as determined in pigs). Copper in milk ceruloplasmin appears to be particularly available for absorption, at least in rats.

Pathways of electron transport to periplasmic nitrate (NapA) and nitrite (NrfA) reductases have been investigated in Campylobacter jejuni, a microaerophilic food-borne pathogen. The nap operon is unusual in lacking napC (encoding a tetra-haem c-type cytochrome) and napF, but contains a novel gene of unknown function, napL. The iron-sulphur protein NapG has a major role in electron transfer to the NapAB complex, but we show that slow nitrate-dependent growth of a napG mutant can be sustained by electron transfer from NrfH, the electron donor to the nitritereductase NrfA. A napL mutant possessed approximately 50% lower NapA activity than the wild type but showed normal growth with nitrate as the electron acceptor. NrfA was constitutive and was shown to play a role in protection against nitrosative stress in addition to the previously identified NO-inducible single domain globin, Cgb. However, nitrite also induced cgb expression in an NssR-dependent manner, suggesting that growth of C. jejuni with nitrite causes nitrosative stress. This was confirmed by lack of growth of cgb and nssR mutants, and slow growth of the nrfA mutant, in media containing nitrite. Thus, NrfA and Cgb together provide C. jejuni with constitutive and inducible components of a robust defence against nitrosative stress.

Among vertebrates able to tolerate periods of oxygen deprivation, the painted and red-eared slider turtles (Chrysemys picta and Trachemys scripta) and the crucian carp (Carassius carassius) are the most extreme and can survive even months of total lack of oxygen during winter. The key to hypoxia...... survival resides in concerted physiological responses, including strong metabolic depression, protection against oxidative damage and – in air breathing animals - redistribution of blood flow. Each of these responses is known to be tightly regulated by nitric oxide (NO) and during hypoxia by its metabolite...... nitrite. The aim of this review is to highlight recent work illustrating the widespread roles of NO and nitrite in the tolerance to extreme oxygen deprivation, in particular in the red-eared slider turtle and crucian carp, but also in diving marine mammals. The emerging picture underscores the importance...

Nitrite uptake into red blood cells (RBCs) precedes its intracellular reactions with hemoglobin (Hb) that forms nitric oxide (NO) during hypoxia. We investigated the uptake of nitrite and its reactions with Hb at different oxygen saturations (So2), using RBCs with (carp and rabbit) and without...... (hagfish and lamprey) anion exchanger-1 (AE1) in the membrane, with the aim to unravel the mechanisms and oxygenation dependencies of nitrite transport. Added nitrite rapidly diffused into the RBCs until equilibrium. The distribution ratio of nitrite across the membrane agreed with that expected from HNO2...... diffusion and AE1-mediated facilitated NO2- diffusion. Participation of HNO2 diffusion was emphasized by rapid transmembrane nitrite equilibration also in the natural AE1 knockouts. Following the equilibration, nitrite was consumed by reacting with Hb, which created a continued inward diffusion controlled...

We evaluated the effect of acute exposure to nitrite on expression of antioxidant and metabolic enzyme genes in gill tissue of advanced juvenile Cherax quadricarinatus. A 48h nitrite exposure was conducted, using four test concentrations (NO2-N=0.5, 1, 1.5 and 2mg L(-1)) plus a control group. The relative mRNA expression of mitochondrial manganese superoxide dismutase (mMnSOD), cytosolic MnSOD (cMnSOD), extracellular copper/zinc SOD (exCu/ZnSOD), catalase (CAT), glutathione S-transferase (GST), arginine kinase (AK), glutamate dehydrogenase (GDH), mitochondrial malate dehydrogenase (mMDH), Na(+)/K(+)-ATPase α-subunit and phosphoenolpyruvate carboxykinase (PEPCK) in gill tissue was measured. Significantly increased mRNA expression was observed for all the antioxidant enzymes after 12 and 24h. After 48h, they all decreased at high nitrite concentrations. The gene expression levels of AK, GDH, mMDH and Na(+)/K(+)-ATPase α-subunit showed similar trends as the antioxidant enzymes. Significant depression of gene expression levels of PEPCK occurred throughout the experimental time at high nitrite concentrations. The results indicated that nitrite could induce oxidative and metabolic stress in C. quadricarinatus, in a time dependent manner, which suggests they could be helpful in predicting sublethal nitrite toxicity and useful in environmental monitoring studies.

Sequential mRNA fluorescence in situ hybridization (mRNA FISH) and fluorescence-assisted cell sorting (SmRFF) was used for the identification of nitrite-reducing bacteria in mixed microbial communities. An oligonucleotide probe labeled with horseradish peroxidase (HRP) was used to target mRNA of nirS, the gene that encodes nitritereductase, the enzyme responsible for the dissimilatory reduction of nitrite to nitric oxide. Clones for nirS expression were constructed and used to provide proof of concept for the SmRFF method. In addition, cells from pure cultures of Pseudomonas stutzeri and denitrifying activated sludge were hybridized with the HRP probe, and tyramide signal amplification was performed, conferring a strongly fluorescent signal to cells containing nirS mRNA. Flow cytometry-assisted cell sorting was used to detect and physically separate two subgroups from a mixed microbial community: non-fluorescent cells and an enrichment of fluorescent, nitrite-reducing cells. Denaturing gradient gel electrophoresis (DGGE) and subsequent sequencing of 16S ribosomal RNA (rRNA) genes were used to compare the fragments amplified from the two sorted subgroups. Sequences from bands isolated from DGGE profiles suggested that the dominant, active nitrite reducers were closely related to Acidovorax BSB421. Furthermore, following mRNA FISH detection of nitrite-reducing bacteria, 16S rRNA FISH was used to detect ammonia-oxidizing and nitrite-oxidizing bacteria on the same activated sludge sample. We believe that the molecular approach described can be useful as a tool to help address the longstanding challenge of linking function to identity in natural and engineered habitats.

Nitrite (NO2-) is a key intermediate in the marine nitrogen (N) cycle. It is produced and consumed throughout the ocean by the dominant processes driving the distribution, availability and speciation of N. However, the accumulation of nitrite is typically confined to depths near the base of the sunlit euphotic zone and in oxygen-deficient zones. These features are known as the primary and secondary nitrite maximum (PNM and SNM), respectively. The processes controlling nitrite accumulation in these features are not fully understood, but are thought to depend on the microbial community composition and its response to environmental conditions. A variety of approaches have been applied to understanding these features since their discovery, with the stable N and oxygen (O) isotope measurements of nitrite being added to this toolkit most recently. Large variations in nitrite N isotope ratios (15N/14N) and dramatic depletions in 15N contrast with more consistent nitrite O isotope ratios (18O/16O) in the SNM. These signals provide unique information about the mechanisms of nitrite consumption in the SNM. By contrast, nitrite in the PNM shows less variation in 15N/14N, but variations in 18O/16O that provide insight into the mechanisms and rates of N cycling there. This review presents a synthesis of nitrite isotope measurements in the marine environment, highlighting the insights that have been gained from these measurements. This article is part of the themed issue 'Biological and climatic impacts of ocean trace element chemistry'.

Abstract The first carboxylate reductase from Trametes versicolor was identified, cloned, and expressed in Escherichia coli. The enzyme reduces aromatic acids such as benzoic acid and derivatives, cinnamic acid, and 3-phenylpropanoic acid, but also aliphatic acids such as octanoic acid are reduced. Graphical abstract

Full Text Available In the genus Azolla rapid abscission of roots from floating fronds occurs within minutes in response to a variety of stresses, including exposure to nitrite. We found that hydrogen peroxide, though itself not an inducer of root abscission, modulates nitrite-induced root abscission by Azolla pinnata in a dose-dependent manner, with 2 mM H2O2 significantly diminishing the responsiveness to 2 mM NaNO2, and 10 mM H2O2 slightly enhancing it. Hypoxia, which has been found in other plants to result in autogenic production of H2O2, dramatically stimulated root abscission of A. pinnata in response to nitrite, especially for plants previously cultivated in medium containing 5 mM KNO3 compared to plants cultivated under N2-fixing conditions without combined nitrogen. Plants, including Azolla, produce the small signaling molecule nitric oxide (NO from nitrite using nitrate reductase. We found Azolla plants to display dose-dependent root abscission in response to the NO donor spermine NONOate. Treatment of plants with the thiol-modifying agents S-methyl methanethiosulfonate or glutathione inhibited the nitrite-induced root abscission response. Synchrotron radiation-based Fourier transform infrared (SR-FTIR spectromicroscopy revealed higher levels of carbonylation in the abscission zone of dropped roots, indicative of reaction products of polysaccharides with potent free radical oxidants. We hypothesize that metabolic products of nitrite and NO react with H2O2 in the apoplast leading to free-radical-mediated cleavage of structural polysaccharides and consequent rapid root abscission.

In the genus Azolla rapid abscission of roots from floating fronds occurs within minutes in response to a variety of stresses, including exposure to nitrite. We found that hydrogen peroxide, though itself not an inducer of root abscission, modulates nitrite-induced root abscission by Azolla pinnata in a dose-dependent manner, with 2 mM H2O2 significantly diminishing the responsiveness to 2 mM NaNO2, and 10 mM H2O2 slightly enhancing it. Hypoxia, which has been found in other plants to result in autogenic production of H2O2, dramatically stimulated root abscission of A. pinnata in response to nitrite, especially for plants previously cultivated in medium containing 5 mM KNO3 compared to plants cultivated under N2-fixing conditions without combined nitrogen. Plants, including Azolla, produce the small signaling molecule nitric oxide (NO) from nitrite using nitrate reductase. We found Azolla plants to display dose-dependent root abscission in response to the NO donor spermine NONOate. Treatment of plants with the thiol-modifying agents S-methyl methanethiosulfonate or glutathione inhibited the nitrite-induced root abscission response. Synchrotron radiation-based Fourier transform infrared spectromicroscopy revealed higher levels of carbonylation in the abscission zone of dropped roots, indicative of reaction products of polysaccharides with potent free radical oxidants. We hypothesize that metabolic products of nitrite and NO react with H2O2 in the apoplast leading to free-radical-mediated cleavage of structural polysaccharides and consequent rapid root abscission.

Literature frequently mentions increased nitrite concentrations along with its inhibitory effect towards bacteria and aquatic life. Nitrite accumulation has been studied for decades, and although numerous causal factors have already been commented on in literature, the mechanism of nitrite

-tolerant species. Nitrite-induced vasodilation is well documented, and many studies support a role of erythrocyte/hemoglobin-derived NO. Vasodilation can, however, also originate from nitrite reduction within the vessel wall, and at present there is no consensus regarding the relative importance of competing...

Inorganic nitrate and nitrite from endogenous or dietary sources are metabolized in vivo to nitric oxide (NO) and other bioactive nitrogen oxides. The nitrate-nitrite-NO pathway is emerging as an important mediator of blood flow regulation, cell signaling, energetics and tissue responses to hypoxia.

Inorganic nitrite (NO2(-), ON-O(-) ←→ (-)O-NO) is the autoxidation product of nitric oxide (NO). Nitrite can also be formed from inorganic nitrate (ONO2(-)), the major oxidation product of NO in erythrocytes, by the catalytic action of bacterial nitrate reductase in gut and oral microflora. Nitrite can be reduced to NO by certain cellular proteins and enzymes, as well as in the gastric juice under acidic conditions. Hemoglobin, xanthine oxidoreductase and carbonic anhydrase (CA) have been reported to convert nitrite to NO. Renal CA isoforms are involved in the reabsorption of nitrite and may, therefore, play an important role in NO homeostasis. Yet, the mechanisms underlying the action of CA on nitrite are incompletely understood. The nitrate/nitrite system is regarded as a reservoir of NO. We have recently shown that nitrite reacts chemically with carbon dioxide (CO2), the regular substrate of CA. The present communication reports a stable isotope ratio mass spectrometry (IRMS) study on the reaction of NO2(-) and CO2 performed in 50 mM HEPES buffer of pH 7.4 at 37 °C. By using (18)O-labelled nitrite ((18)ON-O(-)/(-18)O-NO) and CO2 we observed formation of (18)O-labelled CO2. This finding is an unequivocal evidence of the chemical reaction of (18)ON-O(-)/(-18)O-NO with CO2. The reaction is rapid and involves nucleophilic attack of the negatively charged nitrite via one of its oxygen atoms on the partially positively charged CO2 molecule to form the putative intermediate (18)ON-O-CO2(-)/(-)O2C-(18)O-NO. The by far largest fraction of this intermediate decomposes back to (18)ON-O(-)/(-18)O-NO and CO2. A very small fraction of the intermediate, however, rearranges and finally decomposes to form (18)OCO and nitrite. This reaction is slower in the presence of an isolated erythrocytic CA isoform II. In summary, NO2(-), CO2 and CA are ubiquitous. The chemical reaction of NO2(-) with CO2 and its modulation by CA isoforms may play important roles in the transport of

The effect of nitrite stress induced in Paracoccus denitrificans PD1222 was examined using additions of sodium nitrite to an aerobic bacterial culture. Nitrite generates a strong stress response in P. denitrificans, causing growth inhibition. This is dependent on both the concentration of nitrite present and the pH. The pH dependent effect of nitrite growth inhibition is likely a result of nitrite and free nitrous acid (FNA; pKa = 3.16) and subsequent reactive nitrogen oxides, ...

Nitrate Reductase (NR) is a rating-limit and key enzyme of nitrate assimilation in plants ,so ,NR activity is important for growth,development and the dry matter accumulation of plants. The regulation of NR activity appears to be rather complex and many studies have been devoted to the description of regulation and properties,but in this paper we focus on the properties and regulation of NR in higher plants.

There is an increasing interest in the study of NO chemical affinities of organic nitrites, for the bio-logical and physiological effects of organic nitrites seem to be due to their ability to release NO. In this paper, NO chemical affinities of ten substituted benzyl nitrites were determined by titration calorimetry combined with a ther-modynamic cycle in acetonitrile solution. The results show that ΔHhet(O-NO)s of benzyl nitrites are substan-tially larger than the corresponding ΔHhomo(O-NO)s, suggesting that these O-nitroso compounds much more easily release NO radicals by the O-NO bond homolytic cleavage. It is believed that the structural and energetic information disclosed in this work should be useful in understanding chemical and biological functions of organic nitrites.

A simple spectrophotometric method for the trace determination of nitrite (NO/sub 2//sup /minus//) is described. Nitrite is reacted with Salbutamol sulfate in acidic medium which gives a yellow color in alkaline medium (less than or equal to pH 7) and can be determined in the presence of several cations and anions. Beer's law is obeyed in the range of 1.8 to 27.6 ppm of nitrite with the molar absorptivity 1.8 /times/ 10/sup 3/ 1 /times/ mole /sup /minus/1/ /times/ cm/sup /minus/1/ at 410 nm. The proposed method can also be utilized for the determination of nitrate (NO/sub 3//sup /minus//) after its reduction to nitrite. The method has been applied for the determination of various samples containing traces of nitrite.

The chemical interplay of nitrogen oxides (NO's) with hemoglobin (Hb) has attracted considerable recent attention because of its potential significance in the mechanism of NO-related vasoactivity regulated by Hb. An important theme of this interplay-redox coupling in adducts of heme iron and NO's-has sparked renewed interest in fundamental studies of FeNO(x) coordination complexes. In this Article, we report combined UV-vis and comprehensive electron paramagnetic resonance (EPR) spectroscopic studies that address intriguing questions raised in recent studies of the structure and affinity of the nitrite ligand in complexes with Fe(III) in methemoglobin (metHb). EPR spectra of metHb/NO(2)(-) are found to exhibit a characteristic doubling in their sharper spectral features. Comparative EPR measurements at X- and S-band frequencies, and in D(2)O versus H(2)O, argue against the assignment of this splitting as hyperfine structure. Correlated changes in the EPR spectra with pH enable complete assignment of the spectrum as deriving from the overlap of two low-spin species with g values of 3.018, 2.122, 1.45 and 2.870, 2.304, 1.45 (values for samples at 20 K and pH 7.4 in phosphate-buffered saline). These g values are typical of g values found for other heme proteins with N-coordinated ligands in the binding pocket and are thus suggestive of N-nitro versus O-nitrito coordination. The positions and shapes of the spectral lines vary only slightly with temperature until motional averaging ensues at approximately 150 K. The pattern of motional averaging in the variable-temperature EPR spectra and EPR studies of Fe(III)NO(2)(-)/Fe(II)NO hybrids suggest that one of two species is present in both of the alpha and beta subunits, while the other is exclusive to the beta subunit. Our results also reconfirm that the affinity of nitrite for metHb is of millimolar magnitude, thereby making a direct role for nitrite in physiological hypoxic vasodilation difficult to justify.

The present invention relates to bacterial enzymes, in particular to an acyl-CoA reductase and a gene encoding an acyl-CoA reductase, the amino acid and nucleic acid sequences corresponding to the reductase polypeptide and gene, respectively, and to methods of obtaining such enzymes, amino acid sequences and nucleic acid sequences. The invention also relates to the use of such sequences to provide transgenic host cells capable of producing fatty alcohols and fatty aldehydes.

Copper(II) is known to catalyze the generation of reactive nitrogen species in the presence of hydrogen peroxide, nitrite or nitric oxide, leading to tyrosine nitration, a biomarker for free radical species associated diseases. Here, we find that biological antioxidants such as ascorbic acid can promote tyrosine nitration in the presence of copper(II) and nitrite under aerobic and weak acidic conditions. Tyrosine nitration is demonstrated on both the β-amyloid peptide and angiotensin I. These studies show that (i) ascorbic acid works as a pro-oxidant in the presence of copper(II) to induce oxidation and nitration on peptides, (ii) both free and coordinated copper(II) can catalyze peptide oxidation and nitration, (iii) nitration occurs under mild acidic conditions (pH = 6.0-6.5).

Anaerobic ammonium oxidation (ANAMMOX) technology has potential technical superiority and economical efficiency for the nitrogen removal from landfill leachate, which contains high-strength ammonium nitrogen (NH4+-N) and refractory organics. To complete the ANAMMOX process, a preceding partial nitritation step to produce the appropriate ratio of nitrite/ammonium is a key stage. The objective of this study was to determine the optimal conditions to acquire constant partial nitritation for landfill leachate treatment, and a bench scale fixed bed bio-film reactor was used in this study to investigate the effects of the running factors on the partial nitritation. The results showed that both the dissolved oxygen (DO) concentration and the ammonium volumetric loading rate (Nv) had effects on the partial nitritation. In the controlling conditions with a temperature of 30±1℃, Nv of 0.2-1.0 kg NH4+-N/(m3·d), and DO concentration of 0.8-2.3 mg/L, the steady partial nitritation was achieved as follows: more than 94% partial nitritation efficiency (nitrite as the main product), 60%-74% NH4+-N removal efficiency, and NO2--N/NH4+-N ratio (concentration ratio) of 1.0-1.4 in the effluent.The impact of temperature was related to Nv at certain DO concentration, and the temperature range of 25-30℃ was suitable for treating high strength ammonium leachate. Ammonium-oxidizing bacteria (AOB) could be acclimated to higher FA (free ammonium) in the range of 122-224 mg/L. According to the denaturing gradient gel electrophoresis analysis result of the bio-film in the reactor, there were 25 kinds of 16S rRNA gene fragments, which indicated that abundant microbial communities existed in the bio-film, although high concentrations of ammonium and FA may inhibit the growth of the nitrite-oxidizing bacteria (NOB) and other microorganisms in the reactor.

Colorimetric methods are still important for determining nitrate and nitrite. A critical step in the use of these methods to determine nitrate in low concentrations is the reaction time required to totally reduce nitrate to nitrite, i.e., 24h in the dark. This work involved a study of the influence of ultrasonic irradiation on the nitrate reduction reaction by hydrazine. Our findings indicated that ultrasonic irradiation, associated with copper(II) ion as a catalyst, increased the redox reaction rate, decreasing the reaction time to about 10min when the power of the ultrasonic irradiation was set in 14.0357W. The strong influence of the ultrasonic irradiation in the reduction reaction rates can be sustained by an excellent linear correlation (R(2)=0.9993) between the kinetic constants and ultrasonic powers. Nitrate conversion also increased from 68% to 98% at the latter conditions. It thus become clear that high intensity ultrasound is very beneficial for this reduction reaction to proceed in good yield and in short reaction time in comparison to its silent reaction.

Sodium nitrite has been identified as a key antimicrobial ingredient to control pathogens in ready-to-eat (RTE) meat and poultry products, including Listeria monocytogenes. This study was designed to more clearly elucidate the relationship between chemical factors (ingoing nitrite, ascorbate, and residual nitrite) and L. monocytogenes growth in RTE meats. Treatments of cooked, cured pork sausage (65% moisture, 1.8% salt, pH 6.6, and water activity 0.98) were based on response surface methodology with ingoing nitrite and ascorbate concentrations as the two main factors. Concentrations of nitrite and ascorbate, including star points, ranged from 0 to 352 and 0 to 643 ppm, respectively. At one of two time points after manufacturing (days 0 and 28), half of each treatment was surface inoculated to target 3 log CFU/g of a five-strain L. monocytogenes cocktail, vacuum packaged, and stored at 7°C for up to 4 weeks. Growth of L. monocytogenes was measured twice per week, and enumerations were used to estimate lag time and growth rates for each treatment. Residual nitrite concentrations were measured on days 0, 4, 7, 14, 21, and 28, and nitrite depletion rate was estimated by using first-order kinetics. The response surface methodology was used to model L. monocytogenes lag time and growth rate based on ingoing nitrite, ascorbate, and the residual nitrite remaining at the point of inoculation. Modeling results showed that lag time was impacted by residual nitrite concentration remaining at inoculation, as well as the squared term of ingoing nitrite, whereas growth rate was affected by ingoing nitrite concentration but not by the remaining residual nitrite at the point of inoculation. Residual nitrite depletion rate was dependent upon ingoing nitrite concentration and was only slightly affected by ascorbate concentration. This study confirmed that ingoing nitrite concentration influences L. monocytogenes growth in RTE products, yet residual nitrite concentration contributes

Nitric oxide (NO) is a signal molecule with functions such as neurotransmission, local vascular relaxation, and anti-inflammation in many physiological and pathological processes. Various factors regulate its intracellular lifetime. Due to its high reactivity in biological systems, it is transformed in the bloodstream into nitrates (NO(-)(3)) by oxyhemoglobin. The Griess reaction is a technically simple method (spectrophotometric, 540 nm) for the analysis of nitrites (NO(-)(2)) in aqueous solutions. We studied the interference of common anticoagulants in the quantification of nitrate and nitrite in plasma samples by the Griess method. We obtained rat plasma using heparin or sodium EDTA as anticoagulants, then added, or otherwise, known NO(-)(3) amounts in order to calculate their recovery. We also studied the effect of ultra-filtration performed before Griess reaction on plasma and aqueous solutions of various anticoagulants (heparin, EDTA, and also sodium citrate) to compare the recoveries of added NO(-)(3) or NO(-)(2). We used standards of NO(-)(3) or NO(-)(2) for quantification. We conclude that: (i) The bacterial nitrate reductase used to reduce NO(-)(3) to NO(-)(2) is unstable in certain storage conditions and interferes with different volumes of plasma used. (ii) The ultrafiltration (which is sometimes performed before the Griess reaction) of plasma obtained with EDTA or citrate is not recommended because it leads to overestimation of NO(minus sign)(3). In contrast, ultrafiltration is necessary when heparin is used. (iii) The absorbance at 540 nm attributed to plasma itself (basal value or background) interferes in final quantification, especially when ultrafiltration is not performed. For the quantification of plasma NO(-)(3) we recommend: sodium EDTA as anticoagulant, no ultrafiltration of plasma, and measurement of the absorbance background of each sample.

In catalyzing the reversible hydration of CO2 to bicarbonate and protons, the ubiquitous enzyme carbonic anhydrase (CA) plays a crucial role in CO2 transport, in acid-base balance, and in linking local acidosis to O2 unloading from hemoglobin. Considering the structural similarity between...... in the reaction induces vasodilation in aortic rings. This reaction occurs under normoxic and hypoxic conditions and in various tissues at physiological levels of CA and nitrite. Furthermore, two specific inhibitors of the CO2 hydration, dorzolamide and acetazolamide, increase the CA-catalyzed production...... of vasoactive NO from nitrite. This enhancing effect may explain the known vasodilating effects of these drugs and indicates that CO2 and nitrite bind differently to the enzyme active site. Kinetic analyses show a higher reaction rate at high pH, suggesting that anionic nitrite participates more effectively...

A new fluorescence quenching method has been developed to determine nitrite in water samples. The method is based on the diazo-reaction between nitrite ion and safranine T in hydrochloric acid medium. The fluorescence intensity was measured at excitation and emission wavelengths of 525.0 and 556.0 nm, respectively. The optimum experimental conditions have been confirmed and the concentration of hydrochloric acid was 0.096 mol.L-1; the reaction time was 10 min. Under the selected conditions, the relationship was obtained between the fluorescence intensity and nitrite concentration in the range 4-200 x 10(-9) g.mL-1 NO2-. The detection limit was 4 x 10(-9) g.mL-1 NO2-. The method was applied to determine trace nitrite in water samples. The mechanism involved in the reaction was discussed.

Full Text Available Microbial fuel cell (MFC with nitrite as an electron acceptor in cathode provided a new technology for nitrogen removal and electricity production simultaneously. The influences of influent nitrite concentration and external resistance on the performance of denitrifying MFC were investigated. The optimal effectiveness were obtained with the maximum total nitrogen (TN removal rate of 54.80±0.01 g m-3 d-1. It would be rather desirable for the TN removal than electricity generation at lower external resistance. Denaturing gradient gel electrophoresis suggested that Proteobacteria was the predominant phylum, accounting for 35.72%. Thiobacillus and Afipia might benefit to nitrite removal. The presence of nitrifying Devosia indicated that nitrite was oxidized to nitrate via a biochemical mechanism in the cathode. Ignavibacterium and Anaerolineaceae was found in the cathode as a heterotrophic bacterium with sodium acetate as substrate, which illustrated that sodium acetate in anode was likely permeated through proton exchange membrane to the cathode .

The rumen bacterium Wolinella succinogenes grows by respiratory nitrate ammonification with formate as electron donor. Whereas the enzymology and coupling mechanism of nitrite respiration is well known, nitrate reduction to nitrite has not yet been examined. We report here that intact cells and cell fractions catalyse nitrate and chlorate reduction by reduced viologen dyes with high specific activities. A gene cluster encoding components of a putative periplasmic nitrate reductase system (napA, G, H, B, F, L, D) was sequenced. The napA gene was inactivated by inserting a kanamycin resistance gene cassette. The resulting mutant did not grow by nitrate respiration and did not reduce nitrate during growth by fumarate respiration, in contrast to the wild type. An antigen was detected in wild-type cells using an antiserum raised against the periplasmic nitrate reductase (NapA) from Paracoccus pantotrophus. This antigen was absent in the W. succinogenes napA mutant. It is concluded that the periplasmic nitrate reductase NapA is the only respiratory nitrate reductase in W. succinogenes, although a second nitrate-reducing enzyme is apparently induced in the napA mutant. The nap cluster of W. succinogenes lacks a napC gene whose product is thought to function in quinol oxidation and electron transfer to NapA in other bacteria. The W. succinogenes genome encodes two members of the NapC/NirT family, NrfH and FccC. Characterization of corresponding deletion mutants indicates that neither of these two proteins is required for nitrate respiration. A mutant lacking the genes encoding respiratory nitritereductase (nrfHA) had wild-type properties with respect to nitrate respiration. A model of the electron transport chain of nitrate respiration is proposed in which one or more of the napF, G, H and L gene products mediate electron transport from menaquinol to the periplasmic NapAB complex. Inspection of the W. succinogenes genome sequence suggests that ammonia formation from

Full Text Available Agricultural advancement and population growth have prompted increases in food supplies, and higher crop yields have been made possible through the application of fertilizers. Large quantities of livestock and poultry on farms, along with the accumulation of biomass and agricultural residues, can cause contamination of ground water resources and other water sanitation concerns in both developing and developed countries. Nitrate is mainly used as a fertilizer in agriculture, and because of its high solubility in water, it can create biological problems in the environment. High usage of nitrite in the food industry as a preservative, flavor enhancer, antioxidant, and color stabilizing agent can cause human exposure to this toxic compound. Nitrite is 10 times as toxic as nitrate in humans. Nitrate is converted to nitrite and nitrosamine compounds in the human stomach, which can lead to bladder cancer. In this review, sources of nitrate and nitrite exposure were investigated. Furthermore, the review evaluates standard levels of nitrate and nitrite in different foods, and acceptable daily doses of these compounds in various countries. Finally, we discuss valid methods of nitrate and nitrite identification and removal in foods.

When the AIDS epidemic was in its earliest stages, and prior to identification of HIV as the etiological factor, the use of volatile nitrites by the male homosexual community to enhance sexual activities appeared to have a significant role in this disease. Preliminary observations indicated that that portion of the male homosexual community which developed Kaposi's sarcoma were also heavy nitrite users. These nitrites had been demonstrated to be mutagenic in bacteria and thus it was postulated that they could be responsible for the appearance of the sarcoma. To evaluate further the genotoxic activity of these chemicals, six nitrites, including those most commonly used by homosexuals for sexual gratification, were selected for testing in the mouse lymphoma TK {plus minus} and Salmonell typhimurium mutagenicity assays. One chemical, n-amyl nitrite, was negative in the mouse lymphoma assay, while the other five chemicals, n-butyl, isobutyl, iso-amyl, sec-butyl, and n-propyl nitrite, were positive. All six compounds were positive in the Salmonella assay. The mutagenic and known toxic effects of these chemicals remain a concern because a large population of teenagers and young adults continue to abuse these substances.

The goal of this study was to elucidate the mechanisms of nitrous oxide (N2O) production from a bioreactor for partial nitrification (PN). Ammonia-oxidizing bacteria (AOB) enriched from a sequencing batch reactor (SBR) were subjected to N2O production pathway tests. The N2O pathway test was initiated by supplying an inorganic medium to ensure an initial NH4(+)-N concentration of 160 mg-N/L, followed by (15)NO2(-) (20 mg-N/L) and dual (15)NH2OH (each 17 mg-N/L) spikings to quantify isotopologs of gaseous N2O ((44)N2O, (45)N2O, and (46)N2O). N2O production was boosted by (15)NH2OH spiking, causing exponential increases in mRNA transcription levels of AOB functional genes encoding hydroxylamine oxidoreductase (haoA), nitritereductase (nirK), and nitric oxide reductase (norB) genes. Predominant production of (45)N2O among N2O isotopologs (46% of total produced N2O) indicated that coupling of (15)NH2OH with (14)NO2(-) produced N2O via N-nitrosation hybrid reaction as a predominant pathway. Abiotic hybrid N2O production was also observed in the absence of the AOB-enriched biomass, indicating multiple pathways for N2O production in a PN bioreactor. The additional N2O pathway test, where (15)NH4(+) was spiked into 400 mg-N/L of NO2(-) concentration, confirmed that the hybrid N2O production was a dominant pathway, accounting for approximately 51% of the total N2O production.

Full Text Available Background and Objectives: Sodium nitrite and potassium nitrite have been traditionally used for inhibition of Clostridium botulinum and also as an agent to stabilize the color of meat products; however, usage of these additives at high levels could lead to toxicity and cancer originating from the formation of nitrosamines. Nowadays, application of natural preservatives in order to reduce the nitrite content in meat products is increasing. Thus, we used dry red grape pomace (DRGP as a natural alternative to sodium nitrite. Materials and Methods: The effect of two levels of DRGP (1 and 2% on the proximate composition, microbial counts, pH values and residual nitrite level of the samples formulated with two levels of sodium nitrite (30 and 60 mg/kg, as well as the comparison of these sausages with the blank (nitrite-free and control (full nitrite added samples on the 1rst, 10th, 20th and 30th days of storage at 3-5 °C were evaluated. Results: The results showed that all chemical compositions were in the ranges reported by other researchers, and nitrite was very effective in preventing the microbial growth. Also about 50 % of the ingoing nitrite could be analyzed in the samples after processing. Moreover, the residual nitrite level declined both during the storage of sausage and after the addition of DRGP. Conclusions: The use of DRGP in combination with nitrite for sausages was more effective in keeping the quality and safety of the refrigerated consumer products as indicated by the lower nitrite levels, microbial count and similar composition as compared to the samples treated with nitrite and without nitrite. Keywords: Dry red grape pomace (DRGP, Sausage, Nitrite, Microbial count

The food additive nitrite (E249, E250) is commonly used in meat curing as a food preservation method. Because of potential negative health effects of nitrite, its use is strictly regulated. In an earlier study we have shown that the calculated intake of nitrite in children can exceed the acceptable daily intake (ADI) when conversion from dietary nitrate to nitrite is included. This study examined time-dependent changes in nitrite levels in four Swedish meat products frequently eaten by children: pork/beef sausage, liver paté and two types of chicken sausage, and how the production process, storage and also boiling (e.g., simmering in salted water) and frying affect the initial added nitrite level. The results showed a steep decrease in nitrite level between the point of addition to the product and the first sampling of the product 24 h later. After this time, residual nitrite levels continued to decrease, but much more slowly, until the recommended use-by date. Interestingly, this continuing decrease in nitrite was much smaller in the chicken products than in the pork/beef products. In a pilot study on pork/beef sausage, we found no effects of boiling on residual nitrite levels, but frying decreased nitrite levels by 50%. In scenarios of time-dependent depletion of nitrite using the data obtained for sausages to represent all cured meat products and including conversion from dietary nitrate, calculated nitrite intake in 4-year-old children generally exceeded the ADI. Moreover, the actual intake of nitrite from cured meat is dependent on the type of meat source, with a higher residual nitrite levels in chicken products compared with pork/beef products. This may result in increased nitrite exposure among consumers shifting their consumption pattern of processed meats from red to white meat products.

Copper-exchanged and acidic zeolites are shown to produce nitric oxide (NO) from a nitrite source in biologically active (nanomolar) concentrations. Four zeolites were studied; mordenite, ferrierite, ZSM-5 and SSZ-13, which had varying pore size, channel systems and Si/Al ratios. ZSM-5 and SSZ-13 produced the highest amounts of NO in both the copper and acid form. The high activity and regeneration of the copper active sites makes them good candidates for long-term NO production. Initial cytotoxicity tests have shown at least one of the copper zeolites (Cu-SSZ-13) to be biocompatible, highlighting the potential usage within biomedical applications.

In Sweden and Finland the spent nuclear fuel is planned to be encapsulated in cast iron canisters that have an outer shield made of copper. The copper shield is responsible for the corrosion protection of the canister construction. General corrosion of the copper is not expected to be the limiting factor in the waste repository environment when estimating the life-time of the canister construction. However, different forms of localised corrosion, i.e. pitting, stress corrosion cracking, or environmentally assisted creep fracture may cause premature failure of the copper shield. Of the probable constituents in the groundwater, nitrites, chlorides, sulphides and carbonates have been suggested to promote localised corrosion of copper. The main assumption made in planning this research program is that the surface films forming on copper in the repository environment largely determine the susceptibility of copper to the different forms of localised corrosion. The availability of reactants, which also may become corrosion rate limiting, is investigated in several other research programs. This research program consists of a set of successive projects targeted at characterising the properties of surface films on copper in repository environment containing different detrimental anions. A further aim was to assess the significance of the anion-induced changes in the stability of the oxide films with regard to localised corrosion of copper. This report summarises the results from a series of investigations on properties of surface films forming on copper in water of pH = 8.9 at temperature of 80 deg C and pressure of 2 MPa. The main results gained so far in this research program are as follows: The surface films forming on copper in the thermodynamic stability region of monovalent copper at 80 deg C consist of a bulk part (about 1 mm thick) which is a good ionic and electronic conductor, and an outer, interfacial layer (0.001 - 0.005 mm thick) which shows p-type semiconductor

Nitrate (NO3-) is one of the most harmful contaminants in the groundwater, and it causes various health problems. Bimetallic catalysts, usually palladium (Pd) coupled with secondary metallic catalyst, are found to properly treat nitrate-containing wastewaters; however, the selectivity toward N2 production over ammonia (NH3) production still requires further improvement. Because the N2 selectivity is determined at the nitrite (NO2-) reduction step on the Pd surface, which occurs after NO3- is decomposed into NO2- on the secondary metallic catalyst, we here performed density functional theory (DFT) calculations and experiments to investigate the NO2- reduction pathway on the Pd surface activated by hydrogen. Based on extensive DFT calculations on the relative energetics among ∼100 possible intermediates, we found that NO2- is easily reduced to NO* on the Pd surface, followed by either sequential hydrogenation steps to yield NH3 or a decomposition step to N* and O* (an adsorbate on Pd is denoted using an asterisk). Based on the calculated high migration barrier of N*, we further discussed that the direct combination of two N* to yield N2 is kinetically less favorable than the combination of a highly mobile H* with N* to yield NH3. Instead, the reduction of NO2- in the vicinity of the N* can yield N2O* that can be preferentially transformed into N2 via diverse reaction pathways. Our DFT results suggest that enhancing the likelihood of N* encountering NO2- in the solution phase before combination with surface H* is important for maximizing the N2 selectivity. This is further supported by our experiments on NO2- reduction by Pd/TiO2, showing that both a decreased H2 flow rate and an increased NO2- concentration increased the N2 selectivity (78.6-93.6% and 57.8-90.9%, respectively).

Nitrite (NO(2)-), being a product of metabolism of both nitric oxide (NO(*)) and nitrate (NO(3)-), can accumulate in tissues and regenerate NO() by several mechanisms. The effect of NO(2)- on ischemia/reperfusion injury was also reported. Nevertheless, the mechanisms of intracellular NO(2)- accumulation are poorly understood. We suggested significant role of nitrite penetration through biological membranes in the form of undissociated nitrous acid (HNO(2)). This hypothesis has been tested using large unilamellar phosphatidylcholine liposomes and several spectroscopic techniques. HNO(2) transport across the phospholipid bilayer of liposomes facilitates proton transfer resulting in intraliposomal acidification, which was measured using pH-sensitive probes. NO(2)(-)-mediated intraliposomal acidification was confirmed by EPR spectroscopy using membrane-impermeable pH-sensitive nitroxide, AMC (2,2,5,5-tetramethyl-1-yloxy-2,5-dihydro-1H-imidazol-3-ium-4-yl)-aminomethanesulfonic acid (pK 5.25), and by (31)P NMR spectroscopy using inorganic phosphate (pK 6.9). Nitrite accumulates inside liposomes in concentration exceeding its concentration in the bulk solution, when initial transmembrane pH gradient (alkaline inside) is applied. Intraliposomal accumulation of NO(2)- was observed by direct measurement using chemiluminescence technique. Perfusion of isolated rat hearts with buffer containing 4 microM NO(2)- was performed. The nitrite concentrations in the effluent and in the tissue, measured after 1 min perfusion, were close, supporting fast penetration of the nitrite through the tissue. Measurements of the nitrite/nitrate showed that total concentration of NO(x) in myocardium increased from initial 7.8 to 24.7 microM after nitrite perfusion. Physiological significance of passive transmembrane transport of NO(2)- and its coupling with intraliposomal acidification are discussed.

Increased nitric oxide (NO) formation is mechanistically linked to pathophysiology of the extrahepatic complications of cirrhosis. NO is formed by either enzymatic or non-enzymatic pathways. Enzymatic production is catalyzed by NO synthase (NOS) while entero-salivary circulation of nitrate and nitrite is linked to non-enzymatic formation of NO under acidic pH in the stomach. There is no data on salivary excretion of nitrate and nitrite in cirrhosis. This study was aimed to investigate salivary levels of nitrate and nitrite in a rat model of biliary cirrhosis. Cirrhosis was induced by bile duct ligation (BDL). Four weeks after the operation, submandibular ducts of anesthetized BDL and control rats were cannulated with polyethylene microtube for saliva collection. Assessment of pH, nitrite and nitrate levels was performed in our research. We also investigated NOS expression by real time RT-PCR to estimate eNOS, nNOS and iNOS mRNA levels in the submandibular glands. Salivary pH was significantly lower in BDL rats in comparison to control animals. We also observed a statistically significant increase in salivary levels of nitrite as well as nitrate in BDL rats while there was no elevation in the mRNA expression of nNOS, eNOS, and iNOS in submandibular glands of cirrhotic groups. This indicates that an increased salivary level of nitrite/nitrate is less likely to be linked to increased enzymatic production of NO in the salivary epithelium. It appears that nitrate/nitrite can be transported from the blood stream by submandibular glands and excreted into saliva as entero-salivary circulation, and this mechanism may have been exaggerated during cirrhosis.

Full Text Available An optical chemical sensor for the determination of nitrite based on incorporating methyltrioctylammonium chloride as an anionic exchanger on the triacetylcellulose polymer has been reported. The response of the sensor is based on the redox reaction between nitrite in aqueous solution and iodide adsorbed on sensing membrane using anion exchange phenomena. The sensing membrane reversibly responses to nitrite ion over the range of 6.52×10-6 - 8.70×10-5 mol L-1 with a detection limit of 6.05×10-7 mol L-1 (0.03 µg mL-1 and response time of 6 min. The relative standard deviation for eight replicate measurements of 8.70×10-6 and 4.34×10-5 mol L-1 of nitrite was 4.4 and 2.5 %, respectively. The sensor was successfully applied for determination of nitrite in food, saliva and water samples.

This process of "curing" food is a long practice that dates back thousands of years long before refrigeration or food safety regulations. Today food safety and mass manufacturing are dependent upon safe and effective means to cure and preserve foods including meats. Nitrite remains the most effective curing agent to prevent food spoilage and bacterial contamination. Despite decades of rigorous research on its safety and efficacy as a curing agent, it is still regarded by many as a toxic undesirable food additive. However, research within the biomedical science community has revealed enormous therapeutic benefits of nitrite that is currently being developed as novel therapies for conditions associated with nitric oxide (NO) insufficiency. Much of the same biochemistry that has been understood for decades in the meat industry has been rediscovered in human physiology. This review will highlight the fundamental biochemistry of nitrite in human physiology and highlight the risk benefit evaluation surrounding nitrite in food and meat products. Foods or diets enriched with nitrite can have profound positive health benefits.

In 2 separate incidents, 6 patients were poisoned with hydrogen sulfide (H2S) in sewer gas. In the first incident, mixing acid- and sodium hydroxide-based drain cleaners in a confined space resulted in 4 poisonings and 2 deaths. Three would-be rescuers were seriously poisoned and 1 died. Two survivors had neurological sequelae. Sodium nitrite appeared to have some clinical efficacy in 1 case. The second incident involved 2 patients working on a pump in a sewage pond. A patient lying on a raft close to the pond surface was seriously poisoned; sodium nitrite was clinically efficacious and this patient survived without developing neurological sequelae. Sodium nitrite deserves further clinical study as a potential H2S antidote.

The effects of chitosan characteristics including the degree of deacetylation,molecular weight,particle size,pH pretreatment and immobilization time on the immobilization of nitrite-oxidizing bacteria (NOB) on biopolymeric chitosan were investigated.Nitrite removal efficiency of immobilized NOB depended on the degree of deacetylation,particle size,pH pretreatment on the surface of chitosan and immobilization time.Scanning electron microscope characterization illustrated that the number of NOB cells attached to the surface of chitosan increased with an increment of immobilization time.The optimal condition for NOB immobilization on chitosan was achieved during a 24-hr immobilization period using chitosan with the degree of deacetylation larger than 80％ and various particle size ranges between 1-5 mm at pH 6.5.In general,the NOB immobilized on chitosan flakes has a high potential to remove excess nitrite from wastewater and aquaculture systems.

In the pentobarbitone/urethane anaesthetized rabbit and pentobarbitone anaesthetized cat intratracheal inhalation of amyl nitrite, a non-specific smooth muscle relaxant, caused a vagally dependent depression of patellar reflex and spontaneous (thermal shivering and external intercostal inspiratory) motor activities. A prolonged vagally-independent potentiation succeeded the initial inhibition. The potentiation of intercostal activity may account for the increase in tidal volume produced by amyl nitrite after vagotomy. The patellar reflex potentiation survived thoracolumbar spinal transection but could not be attributed to a direct facilitation of neuromuscular or muscular events. A correlation existed between the relative dependence of fusimotor support of a muscle and its susceptibility to the inhibitory and subsequent facilitatory affects of amyl nitrite inhalation. The diaphragm and interchondral muscles, essentially independent of the autogenetic facilitation provided by muscle spindles, demonstrated neither of these effects. Conversely, the intercostal muscles depend greatly on such facilitation, as do the pectoral and limb muscles during thermal shivering.

In this study, catalytic generation of nitric oxide by a copper(II) complex embedded within a poly(vinyl chloride) matrix in the presence of nitrite (source of nitric oxide) and ascorbic acid (reducing agent) was shown to effectively control the formation and dispersion of nitrifying bacteria biofilms. Amperometric measurements indicated increased and prolonged generation of nitric oxide with the addition of the copper complex when compared to that with nitrite and ascorbic acid alone. The effectiveness of the copper complex-nitrite-ascorbic acid system for biofilm control was quantified using protein analysis, which showed enhanced biofilm suppression when the copper complex was used in comparison to that with nitrite and ascorbic acid treatment alone. Confocal laser scanning microscopy (CLSM) and LIVE/DEAD staining revealed a reduction in cell surface coverage without a loss of viability with the copper complex and up to 5 mM of nitrite and ascorbic acid, suggesting that the nitric oxide generated from the system inhibits proliferation of the cells on surfaces. Induction of nitric oxide production by the copper complex system also triggered the dispersal of pre-established biofilms. However, the addition of a high concentration of nitrite and ascorbic acid to a pre-established biofilm induced bacterial membrane damage and strongly decreased the metabolic activity of planktonic and biofilm cells, as revealed by CLSM with LIVE/DEAD staining and intracellular adenosine triphosphate measurements, respectively. This study highlights the utility of the catalytic generation of nitric oxide for the long-term suppression and removal of nitrifying bacterial biofilms.

Diabetes mellitus type 2 is a syndrome of disordered metabolism with inappropriate hyperglycemia owing to a reduction in the biological effectiveness of insulin. Type 2 diabetes is associated with an impaired nitric oxide (NO) pathway that probably serves as the key link between metabolic disorders and cardiovascular disease. Insulin-mediated translocation of GLUT4 involves the PI3K/Akt kinase signal cascade that results in activation of endothelial NO synthase (eNOS). eNOS is dysfunctional during diabetes. We hypothesize that loss of eNOS-derived NO terminates the signaling cascade and therefore cannot activate GLUT4 translocation and that dietary nitrite may repair this pathway. In this study, we administered 50mg/L sodium nitrite to db/db diabetic mice for 4 weeks. After 4 weeks treatment, the db/db mice experienced less weight gain, improved fasting glucose levels, and reduced insulin levels. Cell culture experiments using CHO-HIRc-myc-GLUT4eGFP cell lines stably expressing insulin receptor and myc-GLUT4eGFP protein, as well as L6 skeletal muscle cells stably expressing rat GLUT4 with a Myc epitope (L6-GLUT4myc), showed that NO, nitrite, and GSNO stimulate GLUT4 translocation independent of insulin, which is inhibited by NEM. Collectively our data suggest that nitrite improves insulin signaling through restoration of NO-dependent nitrosation of GLUT4 signaling translocation. These data suggest that NO-mediated nitrosation of GLUT4 by nitrite or other nitrosating agents is necessary and sufficient for GLUT4 translocation in target tissue. Description of this pathway may justify a high-nitrate/nitrite diet along with the glycemic index to provide a safe and nutritional regimen for the management and treatment of diabetes.

Objective To obtain efficient nitrite degrading lactic acid bacteria by mutagenesis. Methods With the strain D2 isolated in the preliminary work as the original strain, the UV/NTG mutagenesis were used to prepare for screening. Results After mutagenesis with 15 W UV and 0.5 mg/mL nitrosoguanidine for three times, the new strain obtained degradated 91.4% nitrite (200 mg/L) after cultivation for 24 h, increasing by 12.7%compared with the original. With sodium nitrite as the substrate, the activity of nitritereductase enzyme reached 7.7 mmol/L, increasing by 42.9%. The ability of nitrite degradation and the activity of nitritereductase were stable after several generations. Conclusion A high quality nitrite degrading strain stable genetic cha-racteristics was obtained via UV/NTG mutagenesis.%目的：通过诱变方法获得高效降解亚硝酸盐的优良乳酸菌应用于降低腌制品中的亚硝酸盐。方法以前期实验筛选获得的降解亚硝酸盐性能较强乳酸菌 D2作为初始诱变菌株,采用紫外线和亚硝基胍复合诱变,选育高效降解亚硝酸盐的乳酸菌。结果经15 W紫外线和0.5 mg/mL亚硝基胍三轮复合诱变得到一株优良乳酸菌,该菌株24 h降解亚硝酸盐(200 mg/L)降解率为91.4%,较初始菌株提高了12.7%；以亚硝酸钠为底物,其产亚硝酸盐还原酶的比活力为7.7 mmol/L,较诱变前提高42.9%；连续传代培养后降解亚硝酸盐能力和产亚硝酸盐还原酶活力性能稳定。结论通过紫外线和亚硝基胍复合诱变,获得一株遗传稳定性良好的高效降解亚硝酸盐的菌株。

Full Text Available Abstract Background Generalized hypoxic pulmonary vasoconstriction (HPV occurring during exposure to hypoxia is a detrimental process resulting in an increase in lung vascular resistance. Nebulization of sodium nitrite has been shown to inhibit HPV. The aim of this project was to investigate and compare the effects of nebulization of nitrite and different formulations of acidified sodium nitrite on acute HPV. Methods Ex vivo isolated rabbit lungs perfused with erythrocytes in Krebs-Henseleit buffer (adjusted to 10% hematocrit and in vivo anesthetized catheterized rabbits were challenged with periods of hypoxic ventilation alternating with periods of normoxic ventilation. After baseline hypoxic challenges, vehicle, sodium nitrite or acidified sodium nitrite was delivered via nebulization. In the ex vivo model, pulmonary arterial pressure and nitric oxide concentrations in exhaled gas were monitored. Nitrite and nitrite/nitrate were measured in samples of perfusion buffer. Pulmonary arterial pressure, systemic arterial pressure, cardiac output and blood gases were monitored in the in vivo model. Results In the ex vivo model, nitrite nebulization attenuated HPV and increased nitric oxide concentrations in exhaled gas and nitrite concentrations in the perfusate. The acidified forms of sodium nitrite induced higher levels of nitric oxide in exhaled gas and had longer vasodilating effects compared to nitrite alone. All nitrite formulations increased concentrations of circulating nitrite to the same degree. In the in vivo model, inhaled nitrite inhibited HPV, while pulmonary arterial pressure, cardiac output and blood gases were not affected. All nitrite formulations had similar potency to inhibit HPV. The tested concentration of appeared tolerable. Conclusion Nitrite alone and in acidified forms effectively and similarly attenuates HPV. However, acidified nitrite formulations induce a more pronounced increase in nitric oxide exhalation.

Mesophilic crenarchaeota are frequently found in terrestrial and marine habitats worldwide, but despite their considerable abundance the physiology of these as yet uncultivated archaea has remained unknown. From a 1.2 Gb large-insert environmental fosmid library of a calcareous grassland soil, a ...

and 1 sequence from Sample G840. Cluster VIII contained the majority (23 of 26) of the sequences from Sample V400. Many of the se- quences in Cluster VIII were virtually identical to the nirS sequence of the cultivated denitrifier Pseudo- monas... environmental clone, exhibiting close identity to a cul- tured denitrifier species, was also observed in a recent study of the River Colne estuary sediments, in which virtually identical nirS sequences were obtained from a Flavobacterium isolate and from 2 RT...

There is a functional relationship between the heparan sulfate proteoglycan glypican-1 and the amyloid precursor protein (APP) of Alzheimer disease. In wild-type mouse embryonic fibroblasts, expression and processing of the APP is required for endosome-to-nucleus translocation of anhydromannose-containing heparan sulfate released from S-nitrosylated glypican-1 by ascorbate-induced, nitrosothiol-catalyzed deaminative cleavage. In fibroblasts from the transgenic Alzheimer mouse Tg2576, there is increased processing of the APP to amyloid-β peptides. Simultaneously, there is spontaneous formation of anhydromannose-containing heparan sulfate by an unknown mechanism. We have explored the effect of hypoxia on anhydromannose-containing heparan sulfate formation in wild-type and Tg2576 fibroblasts by deconvolution immunofluorescence microscopy and flow cytometry using an anhydromannose-specific monoclonal antibody and by (35)SO4-labeling experiments. Hypoxia prevented ascorbate-induced heparan sulfate release in wild-type fibroblasts, but induced an increased formation of anhydromannose-positive and (35)S-labeled heparan sulfate in Tg2576 fibroblasts. This appeared to be independent of glypican-1 S-nitrosylation as demonstrated by using a monoclonal antibody specific for S-nitrosylated glypican-1. In hypoxic wild-type fibroblasts, addition of nitrite to the medium restored anhydromannose-containing heparan sulfate formation. The increased release of anhydromannose-containing heparan sulfate in hypoxic Tg2576 fibroblasts did not require addition of nitrite. However, it was suppressed by inhibition of the nitritereductase activity of xanthine oxidoreductase/aldehyde oxidase or by inhibition of p38 mitogen-activated protein kinase or by chelation of iron. We propose that normoxic Tg2576 fibroblasts maintain a high level of anhydromannose-containing heparan sulfate production by a stress-activated generation of nitric oxide from endogenous nitrite. This activation is enhanced

The aldo-keto reductases (AKRs) are one of the three enzyme superfamilies that perform oxidoreduction on a wide variety of natural and foreign substrates. A systematic nomenclature for the AKR superfamily was adopted in 1996 and was updated in September 2000 (visit www.med.upenn.edu/akr). Investigators have been diligent in submitting sequences of functional proteins to the Web site. With the new additions, the superfamily contains 114 proteins expressed in prokaryotes and eukaryotes that are distributed over 14 families (AKR1-AKR14). The AKR1 family contains the aldose reductases, the aldehyde reductases, the hydroxysteroid dehydrogenases and steroid 5beta-reductases, and is the largest. Other families of interest include AKR6, which includes potassium channel beta-subunits, and AKR7 the aflatoxin aldehyde reductases. Two new families include AKR13 (yeast aldose reductase) and AKR14 (Escherichia coli aldehyde reductase). Crystal structures of many AKRs and their complexes with ligands are available in the PDB and accessible through the Web site. Each structure has the characteristic (alpha/beta)(8)-barrel motif of the superfamily, a conserved cofactor binding site and a catalytic tetrad, and variable loop structures that define substrate specificity. Although the majority of AKRs are monomeric proteins of about 320 amino acids in length, the AKR2, AKR6 and AKR7 family may form multimers. To expand the nomenclature to accommodate multimers, we recommend that the composition and stoichiometry be listed. For example, AKR7A1:AKR7A4 (1:3) would designate a tetramer of the composition indicated. The current nomenclature is recognized by the Human Genome Project (HUGO) and the Web site provides a link to genomic information including chromosomal localization, gene boundaries, human ESTs and SNPs and much more.

The effect of biokinetics on nitritation was investigated in two biofilm geometries, the Membrane Aerated Biofilm Reactor (MABR) and a conventional biofilm system. A 1D biofilm model was used and evaluated by global sensitivity analysis using the variance based Sobol method. The main focus...... strongly depends on the chosen kinetic parameters of AOB and NOB. The maximum specific growth rates (μmax,AOB and μmax,NOB) had the strongest impact on nitritation efficiency (NE). In comparison, the counter-diffusion geometry yielded more parameter combinations (27.5%) that resulted in high NE than the co...

The interaction of plasma with liquid generates nitrogen species including nitrite (NO− 2). Therefore, the color developing capacity of plasma-treated water (PTW) as a nitrite source for meat curing was investigated in this study. PTW, which is generated by surface dielectric barrier discharge in air, and the increase of plasma treatment time resulted in increase of nitrite concentration in PTW. The PTW used in this study contains 46 ppm nitrite after plasma treatment for 30 min. To evaluate ...

Objective:To determine whether blood nitrite levels are elevated in patients with leptospirosis. Methods: Male patients fulfilling clinical and epidemiological criteria for a diagnosis of leptospirosis were recruited. Those with MAT titre of≥400 together with those seroconverting to a titer of≥200 were included in the analysis. Serum nitrite levels were measured in these patients and age, sex matched healthy controls. Results:Patients from 3 hospitals (n=75) were screened during a 3 month period from 28th June to 3rd September 2009, of whom 20 were eligible for the study. Serum nitrite levels were found to be significantly higher in patients with acute leptospirosis [n=20, (0.359±0.229)μM] compared to controls [(n=13,(0.216±0.051)μM] (P=0.014). A significant correlation was also observed between the MAT titre and the day of illness (r = 0.547; P<0.0001). Conclusions: Serum nitrite levels are higher in patients with acute leptospirosis compared to age and sex matched controls. No correlation could be assessed with severity of illness, as sample size was inadequate to determine this.

This report describes and evaluates four types of nitrate and nitrite destruction and separation technologies that could be used to treat the aqueous, alkaline, nitrate-bearing mixed waste that is generated by the In-Tank Precipitation (ITP) process at the Savannah River Site (SRS). The technologies considered in this report include thermal, hydrothermal, chemical, and electrochemical technologies.

Nov 21, 2011 ... African Journal of Biotechnology Vol. 10(73), pp. ... wastewater can also be treated efficiently for the removal of nitrite in both batch and continuous scales ... the aquaculture industry's rapid growth and is now regarded as the ...

Accurate assessments of exposure to nitrate in drinking water is a crucial part of epidemiological studies investigating long-term adverse human health effects. However, since drinking water nitrate measurements are usually collected for regulatory purposes, assumptions on (1) the intra-distribution system variability and (2) short-term (seasonal) concentration variability have to be made. We assess concentration variability in the distribution system of nitrate, nitrite, and ammonium, and seasonal variability in all Danish public waterworks from 2007 to 2016. Nitrate concentrations at the exit of the waterworks are highly correlated with nitrate concentrations within the distribution net or at the consumers' taps, while nitrite and ammonium concentrations are generally lower within the net compared with the exit of the waterworks due to nitrification. However, nitrification of nitrite and ammonium in the distribution systems only results in a relatively small increase in nitrate concentrations. No seasonal variation for nitrate, nitrite, or ammonium was observed. We conclude that nitrate measurements taken at the exit of the waterworks are suitable to calculate exposures for all consumers connected to that waterworks and that sampling frequencies in the national monitoring programme are sufficient to describe temporal variations in longitudinal studies.

Although alkyl nitrites are used as recreational drugs, there is only little research data regarding their effects on the central nervous system including their neurotoxicity. This study investigated the neurotoxicity of three representative alkyl nitrites (isobutyl nitrite, isoamyl nitrite, and butyl nitrite), and whether it affected learning/memory function and motor coordination in rodents. Morris water maze test was performed in mice after administrating the mice with varying doses of the substances in two different injection schedules of memory acquisition and memory retention. A rota-rod test was then performed in rats. All tested alkyl nitrites lowered the rodents' capacity for learning and memory, as assessed by both the acquisition and retention tests. The results of the rota-rod test showed that isobutyl nitrite in particular impaired motor coordination in chronically treated rats. The mice chronically injected with isoamyl nitrite also showed impaired function, while butyl nitrite had no significant effect. The results of the water maze test suggest that alkyl nitrites may impair learning and memory. Additionally, isoamyl nitrite affected the rodents' motor coordination ability. Collectively, our findings suggest that alkyl nitrites may induce neurotoxicity, especially on the aspect of learning and memory function.

We here discuss the implications of the redox conversion of nitrite to ·NO in the gut, how nitrite-derived ·NO may signal from the digestive to the central nervous system, influencing brain function, as well as a putative ascorbate-driven nitrite/NO pathway occurring in the brain.

Crystals of the oxidized form of the periplasmic nitrate reductase from Cupriavidus necator were obtained using polyethylene glycol 3350 as precipitant The periplasmic nitrate reductase from Cupriavidus necator (also known as Ralstonia eutropha) is a heterodimer that is able to reduce nitrate to nitrite. It comprises a 91 kDa catalytic subunit (NapA) and a 17 kDa subunit (NapB) that is involved in electron transfer. The larger subunit contains a molybdenum active site with a bis-molybdopterin guanine dinucleotide cofactor as well as one [4Fe–4S] cluster, while the small subunit is a di-haem c-type cytochrome. Crystals of the oxidized form of this enzyme were obtained using polyethylene glycol 3350 as precipitant. A single crystal grown at the High Throughput Crystallization Laboratory of the EMBL in Grenoble diffracted to beyond 1.5 Å at the ESRF (ID14-1), which is the highest resolution reported to date for a nitrate reductase. The unit-cell parameters are a = 142.2, b = 82.4, c = 96.8 Å, β = 100.7°, space group C2, and one heterodimer is present per asymmetric unit.

Highlights: • Differences between CuO NP and CuCl{sub 2} exposure were characterized. • Copper accumulation in E. pallida was concentration-dependent. • E. pallida exposed to CuCl{sub 2} accumulated higher copper tissue burdens. • The oxidative stress response was greater in E. pallida exposed to CuO NP. • Both forms of copper inhibited CA activity in E. pallida. - Abstract: Increasing use of metal oxide nanoparticles (NP) by various industries has resulted in substantial output of these NP into aquatic systems. At elevated concentrations, NP may interact with and potentially affect aquatic organisms. Environmental implications of increased NP use are largely unknown, particularly in marine systems. This research investigated and compared the effects of copper oxide (CuO) NP and dissolved copper, as copper chloride (CuCl{sub 2}), on the sea anemone, Exaiptasia pallida. Sea anemones were collected over 21 days and tissue copper accumulation and activities of the enzymes: catalase, glutathione peroxidase, glutathione reductase, and carbonic anhydrase were quantified. The size and shape of CuO NP were observed using a ecanning electron microscope (SEM) and the presence of copper was confirmed by using Oxford energy dispersive spectroscopy systems (EDS/EDX). E. pallida accumulated copper in their tissues in a concentration- and time-dependent manner, with the animals exposed to CuCl{sub 2} accumulating higher tissue copper burdens than those exposed to CuO NP. As a consequence of increased copper exposure, as CuO NP or CuCl{sub 2}, anemones increased activities of all of the antioxidant enzymes measured to some degree, and decreased the activity of carbonic anhydrase. Anemones exposed to CuO NP generally had higher anti-oxidant enzyme activities than those exposed to the same concentrations of CuCl{sub 2}. This study is useful in discerning differences between CuO NP and dissolved copper exposure and the findings have implications for exposure of aquatic

To investigate the correlation between the prophylactic administration of intravenous immunoglobulin (IVIG) to preterm infants and urinary nitrite levels, which can be utilized as an index of endogenous nitric oxide (NO) formation, and to determine if NO formation plays a role in both therapeutic and adverse effects of IVIG. 28 healthy preterm infants were included in this prospective study. They had a mean gestational age of 29.4 +/- 2.2 weeks and weight of 1,387 +/- 371 g. Prophylactic IVIG infusion at a dose of 0.5 g/kg/day was administered when they were 3-10 days old. Urine samples of the neonates were obtained for analysis on days 1, 2 and 3 after IVIG administration as well as 1 day before. Urinary nitrite levels obtained in the subjects were normalized for urinary creatinine concentrations. The mean urinary nitrite levels were: 2.77 +/- 1.66 micromol/mmol creatinine before IVIG administration; 4.33 +/- 3.88 micromol/mmol creatinine on the 1st day of IVIG; 3.77 +/- 2.73 micromol/mmol creatinine on the 2nd day, and 3.64 +/- 3.28 micromol/mmol creatinine on the 3rd day. There was a significant increase in urinary nitrite levels between before and after IVIG administration. There was no statistical difference in urinary nitrate levels between days 1, 2 and 3 after IVIG administration. We demonstrated that urinary nitrite excretion is significantly elevated in preterm infants after prophylactic IVIG administration and this result suggests that endogenous NO formation may play an important role in both the therapeutic and adverse effects of IVIG. Copyright 2000 S. Karger AG, Basel

A system consisting of a two-stage up-flow anaerobic sludge blanket (UASB),an anoxic/aerobic (A/O) reactor and a sequencing batch reactor (SBR),was used to treat landfill leachate.During operation,denitrification and methanogenesis took place simultaneously in the first stage UASB (UASB1),and the effluent chemical oxygen demand (COD) was further removed in the second stage UASB (UASB2).Then the denitrification of nitrite and nitrate in the returned sludge by using the residual COD was accomplished in the A/O reactor,and ammonia was removed via nitrite in it.Last but not least,the residual ammonia was removed in SBR as well as nitrite and nitrate which were produced by nitrification.A system consisting of a two-stage UASB and an A/O reactor was used to achieve the stable short-cut nitrification in the first stage (60 d).The effluent of stage one was treated by SBR in the second stage (60 d).The results over 120 d were as follows: when the total nitrogen (TN) concentration of influent leachate was about 2500 mg/L and the ammonia nitrogen concentration was about 2000 mg/L,the short-cut nitrification with 85%-90% nitrite accumulation was achieved stably in the A/O reactor.The TN and ammonia nitrogen removal efficiencies of the system were 98% and 97%,respectively.The residual ammonia,nitrite and nitrate which were produced by nitrification in the A/O reactor could be washed out almost completely in SBR.The TN and ammonia nitrogen concentrations of final effluent were about 39 mg/L and 12 mg/L,respectively.

The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe–S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism.

A 65-year-old female was investigated due to a gradually increasing greenish colour change of her plastic dental splint, which she used to prevent teeth grinding when sleeping. Furthermore, she had noted a greenish/bluish colour change on the back of her black gloves, which she used to wipe her tears away while walking outdoors. The investigation revealed that the patient had a contact allergy to copper, which is very rare. She had, however, had no occupational exposure to copper. The contact allergy may be caused by long-term exposure of the oral mucosa to copper from copper-rich amalgam fillings, which were frequently used in childhood dentistry up to the 1960s in Sweden. The deposition of a copper-containing coating on the dental splint may be caused by a raised copper intake from drinking water, increasing the copper excretion in saliva, in combination with release of copper due to electrochemical corrosion of dental amalgam. The greenish colour change of the surface of the splint is probably caused by deposition of a mixture of copper compounds, e.g. copper carbonates. Analysis by the X-ray diffraction technique indicates that the dominant component is copper oxide (Cu2O and CuO). The corresponding greenish/bluish discoloration observed on the back of the patient's gloves may be caused by increased copper excretion in tears.

The effect of seawater salinity on nitrite accumulation in short-range nitrification to nitrite as the end productwas studied by using a SBR. Experimental results indicated that the growth of nitrobacteria was inhibited and veryhigh levels of nitrite accumulation at different salinities were achieved under the conditions of 25-28 ℃, pH 7.5-8.0, and the influent ammonia nitrogen of 40-70 mg/L when seawater flow used to flush toilet was less than 35%(salinity 12393 mg/L, Cl- 6778 mg/L) of total domestic wastewater flow, which is mainly ascribed to much highchlorine concentration of seawater. Results showed that high seawater salinity is available for short-range nitrificationto nitrite as the end product. When the seawater flow used to flush toilet accounting for above 70% of the totaldomestic wastewater flow, the removal efficiency of ammonia was still above 80% despite the removal of organicsdeclined obviously(less than 60% ). It was found that the effect of seawater salinity on the removal of organics wasnegative rather than positive one as shown for ammonia removal.

Nitrogen removal via nitrite (the nitrite pathway) is beneficial for carbon-limited biological wastewater treatment plants. However, partial nitrification to nitrite has proven difficult in continuous processes treating domestic wastewater. The nitrite pathway is achieved in this study in a pilot-scale continuous pre-denitrification plant (V=300 L) treating domestic wastewater by controlling the dissolved oxygen (DO) concentration at 0.4-0.7 mg/L. It is demonstrated that the nitrite pathway could be repeatedly and reliably achieved, with over 95% of the oxidized nitrogen compounds at the end of the aerobic zone being nitrite. The nitrite pathway improved the total nitrogen (TN) removal by about 20% in comparison to the nitrate pathway, and also reduced aeration costs by 24%. FISH analysis showed that the nitrite oxidizing bacteria (NOB) population gradually reduced at low DO levels, and reached negligible levels when stable nitrite pathway was established. It is hypothesized that NOB was washed out due to its relatively lower affinity with oxygen. A lag phase was observed in the establishment of the nitrite pathway. Several sludge ages were required for the onset of the nitrite pathway after the application of low DO levels. However, nitrite accumulation increased rapidly after that. A similar lag phase was observed for the upset of the nitrite pathway when a DO concentration of 2-3 mg/L was applied. The nitrite pathway negatively impacted on the sludge settleability. A strong correlation between the sludge volume index and the degree of nitrite accumulation was observed.

Accumulation of lipids and cholesterol by macrophages and subsequent transformation into foam cells are key features in development of atherosclerosis. Serum copper concentrations have been shown to be associated with cardiovascular disease. However, the mechanism behind the proatherogenic effect of copper is not clear. We used DNA microarrays to define the changes in gene expression profile in response to copper exposure of human macrophages. Expression monitoring by DNA microarray revealed 91 genes that were regulated. Copper increased the expression of seven cholesterogenic genes (3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) synthase, IPP isomerase, squalene synthase, squalene epoxidase, methyl sterol oxidase, H105e3 mRNA and sterol-C5-desaturase) and low-density lipoprotein receptor (LDL-R), and decreased the expression of CD36 and lipid binding proteins. The expression of LDL-R and HMG CoA reductase was also investigated using real time PCR. The expression of both of these genes was increased after copper treatment of macrophages (Pmechanism for the association between copper and atherosclerosis. The effect of copper on cholesterogenic genes may also have implications for liver steatosis in early stages of Wilson's disease.

The dramatic growth of copper consumption in China can directly be seen from the expansion of copper products capacity.According to sta- tistics,in the past 4 years,the improvement on the balance of trade on copper bar,copper,and copper alloy and copper wire & cable has driven the growth of copper consumption a lot.

Pinoresinol-lariciresinol and isoflavone reductase classes are phylogenetically related, as is a third, the so-called "isoflavone reductase homologs." This study establishes the first known catalytic function for the latter, as being able to engender the NADPH-dependent reduction of phenylcoumaran benzylic ethers. Accordingly, all three reductase classes are involved in the biosynthesis of important and related phenylpropanoid-derived plant defense compounds. In this investigation, the phenylcoumaran benzylic ether reductase from the gymnosperm, Pinus taeda, was cloned, with the recombinant protein heterologously expressed in Escherichia coli. The purified enzyme reduces the benzylic ether functionalities of both dehydrodiconiferyl alcohol and dihydrodehydrodiconiferyl alcohol, with a higher affinity for the former, as measured by apparent Km and Vmax values and observed kinetic 3H-isotope effects. It abstracts the 4R-hydride of the required NADPH cofactor in a manner analogous to that of the pinoresinol-lariciresinol reductases and isoflavone reductases. A similar catalytic function was observed for the corresponding recombinant reductase whose gene was cloned from the angiosperm, Populus trichocarpa. Interestingly, both pinoresinol-lariciresinol reductases and isoflavone reductases catalyze enantiospecific conversions, whereas the phenylcoumaran benzylic ether reductase only shows regiospecific discrimination. A possible evolutionary relationship among the three reductase classes is proposed, based on the supposition that phenylcoumaran benzylic ether reductases represent the progenitors of pinoresinol-lariciresinol and isoflavone reductases.

Modeling for nitritation process was discussed and analyzed quantlitatively for the factors that influence nitrite accumulation. The results indicated that pH, inorganic carbon source and Hydraulic Retention Time(HRT) as well as biomass concentration are the main factors that influenced the conversion ratio of ammonium to nitrite. A constant high pH can lead to a high nitritation rate and results in high conversion ratio on condition that free ammonia inhibition do not happen. In a CSTR system, without pH control, this conversion ratio can be monitored by pH variation in the reactor. The pH goes down far from the inlet level means a strongly nitrite accumulation. High concentration of alkalinity can promoted the conversion ratio by means of accelerating the nitritation rate through providing sufficient inorganic carbon source(carbon dioxide). When inorganic carbon source was depleted, the nitritation process stopped. HRT adjustment could be an efficient way to make the nitritation system run more flexible, which to some extent can meet the requirements of the fluctuant of inlet parameters such as ammonium concentration, pH, and temperature and so on. Biomass concentration is the key point, especially for a CSTR system in steady state, which was normally circumscribed by the characteristics of bacteria and may also affected by aeration mode and can be increased by prolonging the HRT on the condition of no nitrate accumulation when no recirculation available. The higher the biomass concentration is, the better the nitrite accumulation can be obtained.

In addition to presenting a short history of copper paintings, topics detail artists’ materials and techniques, as well as aspects of the copper industry, including mining, preparation and trade routes.......In addition to presenting a short history of copper paintings, topics detail artists’ materials and techniques, as well as aspects of the copper industry, including mining, preparation and trade routes....

Nitrite (ONO(-)) exerts nitric oxide (NO)-related biological actions and its concentration in the circulation may be of particular importance. Nitrite is excreted in the urine. Hence, the kidney may play an important role in nitrite/NO homeostasis in the vasculature. We investigated a possible involvement of renal carbonic anhydrases (CAs) in endogenous nitrite reabsorption in the proximal tubule. The potent CA inhibitor acetazolamide was administered orally to six healthy volunteers (5 mg/kg) and nitrite was measured in spot urine samples before and after administration. Acetazolamide increased abruptly nitrite excretion in the urine, strongly suggesting that renal CAs are involved in nitrite reabsorption in healthy humans. Additional in vitro experiments support our hypothesis that nitrite reacts with CO(2), analogous to the reaction of peroxynitrite (ONOO(-)) with CO(2), to form acid-labile nitrito carbonate [ONOC(O)O(-)]. We assume that this reaction is catalyzed by CAs and that nitrito carbonate represents the nitrite form that is actively transported into the kidney. The significance of nitrite reabsorption in the kidney and the underlying mechanisms, notably a direct involvement of CAs in the reaction between nitrite and CO(2), remain to be elucidated.

The effect of dissolved oxygen (DO) concentration on nitrite accumulation was investigated in a pilot-scale pre-denitrification process at room temperature for 100 days.In the first 10 days,due to the instability of the system,the DO concentration fluctuated between 1.0 and 2.0 mg/L.In the next 14 days,the DO concentration was kept at 0.5 mg/L and nitrite accumulation occurred,with the average nitrite accumulation rate at 91%.From the 25th day,the DO concentration was increased to 2.0 mg/L to destroy the nitrite accumulation,but nitrite accumulation rate was still as high as 90%.From the 38th day the nitrite accumulation rate decreased to 15%-30% linearly.From the 50th day,DO concentration was decreased to 0.5 mg/L to resume nitrite accumulation.Until the 83rd day the nitrite accumulation rate began to increase to 80%.Dissolved oxygen was the main cause of nitrite accumulation,taking into account other factors such as pH,free ammonia concentration,temperature,and sludge retention time.Because of the different affinity for oxygen between nitrite oxidizing bacteria and ammonia oxidizing bacteria when DO concentration was kept at 0.5 rag/L,nitrite accumulation occurred.

通过基因工程手段增加厌氧氨氧化菌亚硝酸盐还原酶(nitritereductase, nirS)的表达量,运用质粒载体pGEM-T克隆nirS基因。琼脂糖凝胶电泳检测显示, nirS基因重组工程菌在440 bp处有明显的目的条带； nirS基因重组工程菌扩大培养7~8h后即达到生长曲线稳定期,引入外加氮源后,菌体生长情况更优。通过不同菌液投加量以及处理不同初始浓度的亚硝酸钠溶液,检测nirS基因重组工程菌的性能。结果表明,当nirS基因重组工程菌投加30 mL(细菌数为2.3×107个∕mL),亚硝酸盐初始质量浓度为40 mg∕L时,亚硝酸盐去除率达到90%以上。nirS基因重组工程菌可适用于亚硝酸盐废水的处理。%In order to improve the expression quantity of nitritereductase (nirS) in ANAMMOX bacteria through bioengineering means, nirS gene was cloned using the plasmid vector pGEM-T. A target band of 440 bp PCR products from the recombinant genetic engineering bacter was observed by agarose gel electrophoresis. The nirS recombinant genetic engineering bacteria reached stationary phase after 7-8 hours incubation, the addition of nitrogen source was advantageous to the growth of bacteria significantly. The performance of nirS recombinant genetic engineering bacteria was tested by adding different dosages of bacteria and treating sodium nitrite solu‐tion with different initial concentrations. The results showed that, when 30 mL of nirS recombinant genetic engi‐neering bacteria(2.3 × 107 cells/mL) inoculates was added to the solution with 40 mg/L of initial mass concentra‐tion of nitrite, the removal rate of nitrite reached above 90%. It was indicated that nirS recombinant genetic en‐gineering bacteria could be applied for nitrite-containing wastewater treatment in the future.

Copper is a transition metal that has been linked to pathological and beneficial effects in neurodegenerative diseases. In Parkinson's disease, free copper is related to increased oxidative stress, alpha-synuclein oligomerization, and Lewy body formation. Decreased copper along with increased iron has been found in substantia nigra and caudate nucleus of Parkinson's disease patients. Copper influences iron content in the brain through ferroxidase ceruloplasmin activity; therefore decreased protein-bound copper in brain may enhance iron accumulation and the associated oxidative stress. The function of other copper-binding proteins such as Cu/Zn-SOD and metallothioneins is also beneficial to prevent neurodegeneration. Copper may regulate neurotransmission since it is released after neuronal stimulus and the metal is able to modulate the function of NMDA and GABA A receptors. Some of the proteins involved in copper transport are the transporters CTR1, ATP7A, and ATP7B and the chaperone ATOX1. There is limited information about the role of those biomolecules in the pathophysiology of Parkinson's disease; for instance, it is known that CTR1 is decreased in substantia nigra pars compacta in Parkinson's disease and that a mutation in ATP7B could be associated with Parkinson's disease. Regarding copper-related therapies, copper supplementation can represent a plausible alternative, while copper chelation may even aggravate the pathology.

Nitric oxide has been proposed to mediate cytotoxic effects in inflammatory diseases. To investigate the possibility that overproduction of nitric oxide might play a role in the neuropathology of inflammatory and noninflammatory neurological diseases, we compared levels of the markers of nitric oxide, nitrite plus nitrate, in the CSF of controls with those in patients with various neurologic diseases, including Huntington's and Alzheimer's disease, amyotrophic lateral sclerosis, and HIV infection. We found that there were no significant increases in the CSF levels of these nitric oxide metabolites, even in patients infected with HIV or in monkeys infected with poliovirus, both of which have significantly elevated levels of the neurotoxin quinolinic acid and the marker of macrophage activation, neopterin. However, CSF quinolinic acid, neopterin, and nitrite/nitrate levels were significantly increased in a small group of patients with bacterial and viral meningitis.

Full Text Available The acute toxicity study of sodium nitrite in the period of observation for 14 days has been carried out on adult male and female rats. The substance was administered orally iu the form of solution in distilled water. Each animal was observed carefully for six hours after administration, continued twice daily for 1.4 days, those including general appearance, body weight changes, signs of toxicity mortality and gross pathology finding. The signs of toxicity were tremor, ataxia and convulsion; mortalities were occurred in about two hours after administration. Some animal recovered after two - four hours. The LD50 in male mice was 121 (86,6-169,4 mg/kg b.w., in female was 116 (86-155.4 mg/kg b.w. Based on the LD50 value, sodium nitrite is classified as moderately toxic substances.

Nitrate reduction is considered to be a minor microbial pathway in the oxidation of mangrove-derived organic matter due to a limited supply of nitrate in mangrove soils. At a limited availability of this electron acceptor compared to the supply of degradable carbon, nitrate ammonification is thought to be the preferential pathway of nitrate reduction. Mangrove forest mutually differ in their productivity, which may lead to different available carbon to nitrate ratios in their soil. Hence, nitrate ammonification is expected to be of more importance in high- compared to low-productive forests. The hypothesis was tested in flow-through reactors that contain undisturbed mangrove soils from high-productive Avicennia germinans and Rhizophora mangle forests in Florida and low-productive Avicennia marina forests in Saudi Arabia. Nitrate was undetectable in the soils from both regions. It was assumed that a legacy of nitrate ammonification would be reflected by a higher ammonium production from these soils upon the addition of nitrate. Unexpectedly, the soils from the low-productive forests in Saudi Arabia produced considerably more ammonium than the soils from the high-productive forests in Florida. Hence, other environmental factors than productivity must govern the selection of nitrate ammonification or denitrification. A rather intriguing observation was the 1:1 production of nitrite and ammonium during the consumption of nitrate, more or less independent from sampling region, location, sampling depth, mangrove species and from the absence or presence of additional degradable carbon. This 1:1 ratio points to a coupled production of ammonium and nitrite by one group of nitrate-reducing microorganisms. Such a production of nitrite will be hidden by the presence of active nitrite-reducing microorganisms under the nitrate-limited conditions of most mangrove forest soils.

Nitrate reduction is considered to be a minor microbial pathway in the oxidation of mangrove-derived organic matter due to a limited supply of nitrate in mangrove soils. At a limited availability of this electron acceptor compared to the supply of degradable carbon, nitrate ammonification is thought to be the preferential pathway of nitrate reduction. Mangrove forest mutually differ in their productivity, which may lead to different available carbon to nitrate ratios in their soil. Hence, nitrate ammonification is expected to be of more importance in high- compared to low-productive forests. The hypothesis was tested in flow-through reactors that contain undisturbed mangrove soils from high-productive Avicennia germinans and Rhizophora mangle forests in Florida and low-productive Avicennia marina forests in Saudi Arabia. Nitrate was undetectable in the soils from both regions. It was assumed that a legacy of nitrate ammonification would be reflected by a higher ammonium production from these soils upon the addition of nitrate. Unexpectedly, the soils from the low-productive forests in Saudi Arabia produced considerably more ammonium than the soils from the high-productive forests in Florida. Hence, other environmental factors than productivity must govern the selection of nitrate ammonification or denitrification. A rather intriguing observation was the 1:1 production of nitrite and ammonium during the consumption of nitrate, more or less independent from sampling region, location, sampling depth, mangrove species and from the absence or presence of additional degradable carbon. This 1:1 ratio points to a coupled production of ammonium and nitrite by one group of nitrate-reducing microorganisms. Such a production of nitrite will be hidden by the presence of active nitrite-reducing microorganisms under the nitrate-limited conditions of most mangrove forest soils.

Full Text Available Introduction: Ham is a product highly consumed by society; however it contains some elements that make it a non-recommended food. Thus, it has been attempted to eliminate or reduce those components.Material and Methods: Content of sodium, nitrites, peroxides and total coliforms were compared after processes of Washing (W and Simple Cooking (SC in turkey and pork ham in a cross-sectional analytical experimental study. Furthermore, sensory acceptance of thesamples through an acceptance test of five points was evaluated. One-way ANOVA with post hoc Bonferroni were used to assess the mean difference between groups. The value of p≤0.05 was considered statistically significant.Results: Both processes reduced the amount of sodium in both types of ham in statistically significant way (p≤0.001. The major percentage of reduction was presented with SC: 73.4% for turkey ham and 63.5% for pork ham. Likewise, the higher percentage of nitrite reduction was 50.6% in pork ham with SC technique (p≤0.05. Peroxide index decreased with both techniques in both hams without statistical significance. Total coliforms count was kept constant in all samples. Higher sensory acceptance of W concerning SC in all sensory characteristics was observed.Conclusions: W and SC techniques reduce sodium, nitrite and peroxide index without affecting the sanitation of hams. Furthermore, W is accepted in all sensory categories. We recommend using W technique before consumption of the product.

High energy X-ray diffraction has been combined with containerless techniques to determine the structure of a series of alkali and ammonium nitrate and nitrite liquids. The systems have been modelled using molecular dynamics simulation which allows for the flexibility of, and movement of charge within, the molecular anions. The model reproduces the experimentally-determined scattering functions in both the low-and high-Q regimes reflecting the inter-and intra-molecular length-scales. For ammonium nitrate the best fit to the diffraction data is obtained by assuming the NH4+ cation to have a radius closer to that for Cs+ rather than a smaller cation such as Rb+ as often previously assumed. The alkali nitrites show an emergent length scale, attributed to the nitrogen-nitrogen spatial correlations, that depends on both temperature and the identity of the alkali cation. The corresponding nitrates show a more subtle effect in the nitrogen-nitrogen correlations. As a result, the nature of this N-N length-scale appears different for the respective nitrites and nitrates.

Full Text Available Liver ischemia-reperfusion injury is a major cause of primary graft non-function or initial function failure post-transplantation. In this study, we examined the effects of sodium nitrite supplementation on liver IRI in either Lactated Ringer's (LR solution or University of Wisconsin (UW solution. The syngeneic recipients of liver grafts were also treated with or without nitrite by intra-peritoneal injection. Liver AST and LDH release were significantly reduced in both nitrite-supplemented LR and UW preservation solutions compared to their controls. The protective effect of nitrite was more efficacious with longer cold preservation times. Liver histological examination demonstrated better preserved morphology and architecture with nitrite treatment. Hepatocellular apoptosis was significantly reduced in the nitrite-treated livers compared their controls. Moreover, liver grafts with extended cold preservation time of 12 to 24 hours demonstrated improved liver tissue histology and function post-reperfusion with either the nitrite-supplemented preservation solution or in nitrite-treated recipients. Interestingly, combined treatment of both the liver graft and recipient did not confer protection. Thus, nitrite treatment affords significant protection from cold ischemic and reperfusion injury to donor livers and improves liver graft acute function post-transplantation. The results from this study further support the potential for nitrite therapy to mitigate ischemia-reperfusion injury in solid organ transplantation.

Nitrite, in equilibrium with free nitrous acid (FNA), can inhibit both aerobic and anaerobic growth of microbial communities through bactericidal activities that have considerable potential for control of microbial growth in a range of water systems. There has been much focus on the effect of nitrite/FNA on anaerobic metabolism and so, to enhance understanding of the metabolic impact of nitrite/FNA on aerobic metabolism, a study was undertaken with a model denitrifying bacterium Paracoccus denitrificans PD1222. Extracellular nitrite inhibits aerobic growth of P. denitrificans in a pH-dependent manner that is likely to be a result of both nitrite and free nitrous acid (pKa = 3.25) and subsequent reactive nitrogen oxides generated from the intracellular passage of FNA into P. denitrificans. Increased expression of a gene encoding a flavohemoglobin protein (Fhp) (Pden_1689) was observed in response to extracellular nitrite. Construction and analysis of a deletion mutant established Fhp to be involved in endowing nitrite/FNA resistance at high extracellular nitrite concentrations. Global transcriptional analysis confirmed nitrite-dependent expression of fhp and indicated that P. denitrificans expressed a number of stress response systems associated with protein, DNA and lipid repair. It is therefore suggested that nitrite causes a pH-dependent stress response that is due to the production of associated reactive nitrogen species, such as nitric oxide from the internalisation of FNA.

Glutathione reductase (NAD(P)h:oxidized glutathione oxidoreductase, EC 1.6.4.2) has been purified 1000-fold from the cytoplasmic fraction of human platelets. Salts, including the heretofore unreported effect of sodium citrate, activate the NADPH-dependent reduction of oxidized glutathione. Sodium citrate and monovalent salt activation appears to involve multiple sites having different binding affinities. At sub-saturating sodium phosphate, non-linear double reciprocal plots indicative of substrate activation by oxidized glutathione were observed. Initial velocity double reciprocal plots at sub-saturating and saturating concentrations of phosphate generate a family of converging lines. NADP+ is a partial inhibitor, indicating that the reduction of oxidized glutathione can proceed by more than one pathway. FMN, FAD, and riboflavin inhibit platelet glutathione reductase by influencing only the V while nitrofurantoin inhibition is associated with an increase Koxidized glutathione and a decreased V.

The use of nitrite (NaNO2) in the manufacture of salted (cured) meat products has a long tradition in the industry, dating back to the early twentieth century. Nitrite serves several technological purposes, primarily by the formation of a stable red colour in the meat and the inhibition....... Nitrite also has a desirable anti-oxidant activity and contributes to the formation of pleasant flavours. A systematic literature review on the function and use of nitrite in meat leads to a tentative first conclusion that if the level of nitrite added to meat products is sufficient to protect against...... number of experiments were conducted in Denmark in collaboration with the Danish meat manufacturing industry in 1981-1983. Wiltshire bacon and certain canned products largely for export were not investigated in this study, however. The adverse effects of nitrite can mainly be ascribed to the risk...

Striped catfish (Pangasianodon hypophthalmus) are farmed intensively at high stocking densities in Vietnam where they are likely to encounter environmental hypercapnia as well as occasional high levels of aquatic nitrite. Nitrite competes with Cl- for uptake at the branchial HCO3-/Cl- exchanger...... in the initial phase of acid-base compensation. Overall, longer term environmental hypercapnia does not protect against nitrite uptake in P. hypophthalmus, but instead enhances it. In addition, we observed a significant size effect in nitrite accumulation, where large fish attained plasma [nitrite] above...... the ambient concentration, while small fish did not. Small P. hypophthalmus instead had significantly higher plasma [nitrate], and haemoglobin concentrations, revealing greater capacity for detoxifying nitrite by oxidising it to nitrate....

Bacterial genes for the degradation of major dilignols produced in lignifying xylem are expected to be useful tools for the structural modification of lignin in plants. For this purpose, we isolated pinZ involved in the conversion of pinoresinol from Sphingobium sp. strain SYK-6. pinZ showed 43-77% identity at amino acid level with bacterial NmrA-like proteins of unknown function, a subgroup of atypical short chain dehydrogenases/reductases, but revealed only 15-21% identity with plant pinoresinol/lariciresinol reductases. PinZ completely converted racemic pinoresinol to lariciresinol, showing a specific activity of 46±3 U/mg in the presence of NADPH at 30°C. In contrast, the activity for lariciresinol was negligible. This substrate preference is similar to a pinoresinol reductase, AtPrR1, of Arabidopsis thaliana; however, the specific activity of PinZ toward (±)-pinoresinol was significantly higher than that of AtPrR1. The role of pinZ and a pinZ ortholog of Novosphingobium aromaticivorans DSM 12444 were also characterized.

A new tetraruthenated copper(II)-tetra(3,4-pyridyl)porphyrazine species, [CuTRPyPz]{sup 4+}, has been synthesized and fully characterized by means of analytical, spectroscopic and electrochemical techniques. This {pi}-conjugated system contrasts with the related meso-tetrapyridylporphyrins by exhibiting strong electronic interaction between the coordinated peripheral complexes and the central ring. Based on favorable {pi}-stacking and electrostatic interactions, layer-by-layer assembled films were successfully generated from the appropriate combination of [CuTRPyPz]{sup 4+} with copper(II)-tetrasulfonated phtalocyanine, [CuTSPc]{sup 4-}. Their conducting and electrocatalytic properties were investigated by means of impedance spectroscopy and rotating disc voltammetry, exhibiting metallic behavior near the Ru(III/II) redox potential, as well as enhanced catalytic activity for the oxidation of nitrite and sulphite ions. (author)

Understanding nitrite dynamics in oxygen minimum zones (OMZs) is a challenge as it represents an intermediary nitrogen species with a short turnover time. Nitrite is also reduced to nitrogen in OMZs, preventing its accumulation. This creates difficulties in detecting nitrite with colorimetric methods as concentrations may occur below detection limits in some regions. Nitrite concentrations are key to understanding intermediate nitrogen processes and their implication for nitrogen loss in OMZs. A coupled physical-biogeochemical model is applied in the Benguela OMZ to study nitrite dynamics and its associated feedback processes. Simulated results show occurrence of primary and secondary nitrite maxima in the Benguela shelf waters. The primary nitrite maxima in the Benguela are attributed to nitrification and nitrate assimilation as they occur in association with the nitracline. Secondary nitrite maxima accumulate in the Angola-Benguela Front (ABF) OMZ and are attributed to denitrification. The secondary nitrite maxima are consumed by anaerobic ammonium oxidation (anammox) off Walvis Bay. Nitrite maxima are restricted to the shelf off Walvis Bay and advected offshore in the ABF region. Interchanges between the poleward South Atlantic Central Water (SACW) and the equatorward, well-aerated Eastern South Atlantic Central Water (ESACW) drive the seasonality of nitrogen processes in the Benguela. Subsequent nitrite reduction in the Benguela OMZ leads to nitrous oxide production, with high concentrations occurring in the ABF region as a result of nitrification and denitrification. Off Walvis Bay, nitrous oxide production is low since nitrite is consumed by anammox. Nitrous oxide production occurs in thermocline, intermediate and deeper water masses in the ABF region. High N fluxes in the Benguela are attributed to nitrification as compared to anammox and denitrification. Results from this study demonstrate the role of intermediate nitrogen species in nitrogen feedback

The biological nitrogen cycle involves step-wise reduction of nitrogen oxides to ammonium salts and oxidation of ammonia back to nitrites and nitrates by plants and bacteria. Neither process has been thought to have relevance to mammalian physiology; however in recent years the salivary bacterial reduction of nitrate to nitrite has been recognized as an important metabolic conversion in humans. Several enteric bacteria have also shown the ability of catalytic reduction of nitrate to ammonia via nitrite during dissimilatory respiration; however, the importance of this pathway in bacterial species colonizing the human intestine has been little studied. We measured nitrite, nitric oxide (NO) and ammonia formation in cultures of Escherichia coli, Lactobacillus and Bifidobacterium species grown at different sodium nitrate concentrations and oxygen levels. We found that the presence of 5 mM nitrate provided a growth benefit and induced both nitrite and ammonia generation in E.coli and L.plantarum bacteria grown at oxygen concentrations compatible with the content in the gastrointestinal tract. Nitrite and ammonia accumulated in the growth medium when at least 2.5 mM nitrate was present. Time-course curves suggest that nitrate is first converted to nitrite and subsequently to ammonia. Strains of L.rhamnosus, L.acidophilus and B.longum infantis grown with nitrate produced minor changes in nitrite or ammonia levels in the cultures. However, when supplied with exogenous nitrite, NO gas was readily produced independently of added nitrate. Bacterial production of lactic acid causes medium acidification that in turn generates NO by non-enzymatic nitrite reduction. In contrast, nitrite was converted to NO by E.coli cultures even at neutral pH. We suggest that the bacterial nitrate reduction to ammonia, as well as the related NO formation in the gut, could be an important aspect of the overall mammalian nitrate/nitrite/NO metabolism and is yet another way in which the microbiome

Double-chamber microbial fuel cell was applied to investigate the performance of the electricity production and nitrite denitrification through feeding nitrite into the cathode. Factors influencing denitrification performance and power production, such as external resistance, influent nitrite concentration and Nitrite Oxygen Bacteria inhibitors, were studied. The results show that when the concentration of nitrite nitrogen and external resistance were 100 mg L(-1) and 10 Ω, respectively, the nitrite denitrification reached the best state. The NaN3 can inhibit nitrite oxidation effectively; meanwhile, the nitrite denitrification with N2O as the final products was largely improved. The [Formula: see text] was reduced to [Formula: see text], causing the cathode denitrification coulombic efficiency to exceed 100%. In chemoautotrophic bio-nitrification, microorganisms may utilize H2O to oxidize nitrite under anaerobic conditions. Proteobacteria might play a major role in the process of denitrification in MFC.

Full Text Available The effect of nitrite ions on the macrocell corrosion behavior of reinforcing steel embedded in cement mortar was investigated by comparing and analyzing the macrocell corrosion current, macrocell polarization ratios, and slopes of anodic and cathodic steels. Based on the experimental results, the relationship between macrocell potential difference and macrocell current density was analyzed, and the mechanism of macrocell corrosion affected by nitrite ions was proposed. The results indicated that nitrite ions had significant impact on the macrocell polarization ratios of cathode and anode. The presence of nitrite could reduce the macrocell current by decreasing the macrocell potential difference and increasing the macrocell polarization resistance of the anode.

The uptake, depuration and toxicity of environmental nitrite was characterized in Litopenaeus vannamei exposed in water containing low concentrations of artificial sea salt or mixed salts. In 2 g/L artificial sea salts, nitrite was concentrated in the hemolymph in a dose-dependent and rapid manner (steady-state in about 2 d). When exposed to nitrite in 2 g/L artificial sea salts for 4 d and then moved to a similar environment without added nitrite, complete depuration occurred within a day. Increasing salinity up to 10 g/L decreased uptake of environmental nitrite. Nitrite uptake in environments containing 2 g/L mixed salts (combination of sodium, potassium, calcium and magnesium chlorides) was similar to or lower than rates in 2 g/L artificial sea salt. Toxicity was inversely related to total dissolved salt and chloride concentrations and was highest in 2 g/L artificial sea salt (96-h medial lethal concentration = 8.4 mg/L nitrite-N). Animals that molted during the experiments did not appear to be more susceptible to nitrite than animals that did not molt. The shallow slope of the curve describing the relationship between toxicity and salinity suggests that management of nitrite toxicity in low-salinity shrimp ponds by addition of more salts may not be practical. ?? Copyright by the World Aquaculture Society 2004.

Recent evidence from humans and rats indicates that nitrite is a vasodilator under hypoxic conditions by reacting with metal-containing proteins to produce nitric oxide (NO). We tested the hypothesis that near-physiological concentrations of nitrite would produce vasodilation in a hypoxia- and concentration-dependent manner in the hind limb of sheep. Anesthetized sheep were instrumented to measure arterial blood pressure and femoral blood flows continuously in both hind limbs. Nitrite was infused into one femoral artery to raise the nitrite concentration in the femoral vein by 10 to 15-fold while the sheep breathed 50%, 14% or 12% oxygen in inspired air. In contrast to reports in humans and rats, the nitrite infusion had no measurable effect on mean femoral blood flows or vascular conductances, regardless of inspired O2 levels. In vitro experiments showed no significant difference in the release of NO from nitrite in sheep and human red blood cells. Further experiments demonstrated nitrite is converted to NO in rat artery homogenates faster than sheep arteries, and that this source of NO production is attenuated in the presence of a heme oxidizer. Finally, western blots indicate that concentrations of the heme-containing protein cytoglobin, but not myoglobin, are markedly lower in sheep arteries compared with rats. Overall, the results demonstrate that nitrite is not a physiological vasodilator in sheep. This is likely due to a lack of conversion of nitrite to NO within the vascular smooth muscle, perhaps due to deficient amounts of the heme-containing protein cytoglobin.

Full Text Available This paper represents the analytical data of nitrite level obtained from the experimental work done on meat processed samples taken from a meat processing plant in Tirana. There has been a long debate and health concern about the nitrite content in meat products. Nitrite is added to e.g. sausages, and hams and other meat products to preserve these products and keep them free from dangerous bacteria. Among the aims are preventing botulism, a dangerous food poison. But also it’s important to use the smallest possible amount of nitrite as a preservative because nitrite in meat can also form nitrosamines, which can damage the health. That’s why the role of nitrite in processed meat and its recommended level conform to new EC Regulations are given in the introduction part of this paper. It is important that the nitrite level be monitored during all the processing steps up to the end consumers. This makes the objective of this paper. It gives the analytical data on nitrite level on meat processed samples taken and tested during their storage and ripening period of time. Different kinds of meat products are taken and tested to evaluate the influence of various parameters (storage time, time until to the end consumers, various kinds of packing in the degradation rate of ingoing nitrite.

Full Text Available The study relates to determination of nitrate in presence of nitrite in water and can be used in the quality monitoring of natural water (surface and groundwater, drinking water, water from fish farms and public aquaria where autonomous filters is used. The nature and quantity of reagents used have insignificant impact on natural waters and sewages. According to the investigation, the method includes the removal of nitrite from the solution/water with sulfaminic acid, the nitrate ion reduction to nitrite using a reducing mixture that contains Na2SO4 and zinc dust in ratio of 100:5 and determining the nitrite with the Griess reagent.

Copper is an essential dietary component, being the coenzyme of many enzymes with oxidase activity, e.g. ceruloplasmin, superoxide dismutase, monoamine oxidase, etc. The metabolism of copper is complex and imperfectly known. Active transport of copper through the intestinal epithelial cells involves metallothionein, a protein rich in sulfhydryl groups which also binds the copper in excess and probably prevents absorption in toxic amounts. In hepatocytes a metallothionein facilitates absorption by a similar mechanism and regulates copper distribution in the liver: incorporation in an apoceruloplasmin, storage and synthesis of copper-dependent enzymes. Metallothioneins and ceruloplasmin are essential to adequate copper homeostasis. Apart from genetic disorders, diseases involving copper usually result from hypercupraemia of varied origin. Wilson's disease and Menkes' disease, although clinically and pathogenetically different, are both marked by low ceruloplasmin and copper serum levels. The excessive liver retention of copper in Wilson's disease might be due to increased avidity of hepatic metallothioneins for copper and decreased biliary excretion through lysosomal dysfunction. Menkes' disease might be due to low avidity of intestinal and hepatic metallothioneins for copper. The basic biochemical defect responsible for these two hereditary conditions has not yet been fully elucidated.

Full Text Available Nitrate reduction is considered to be a minor microbial pathway in the oxidation of mangrove-derived organic matter due to a limited supply of nitrate in mangrove soils. At a limited availability of this electron acceptor compared to the supply of degradable carbon, nitrate ammonification is thought to be the preferential pathway of nitrate reduction. Mangrove forest mutually differ in their productivity, which may lead to different available carbon to nitrate ratios in their soil. Hence, nitrate ammonification is expected to be of more importance in high- compared to low-productive forests.The hypothesis was tested in flow-through reactors that contain undisturbed mangrove soils from high-productive Avicennia germinans and Rhizophora mangle forests in Florida and low-productive Avicennia marina forests in Saudi Arabia. Nitrate was undetectable in the soils from both regions. It was assumed that a legacy of nitrate ammonification would be reflected by a higher ammonium production from these soils upon the addition of nitrate. Unexpectedly, the soils from the low-productive forests in Saudi Arabia produced considerably more ammonium than the soils from the high-productive forests in Florida. Hence, other environmental factors than productivity must govern the selection of nitrate ammonification or denitrification. A rather intriguing observation was the 1:1 production of nitrite and ammonium during the consumption of nitrate, more or less independent from sampling region, location, sampling depth, mangrove species and from the absence or presence of additional degradable carbon. This 1:1 ratio points to a coupled production of ammonium and nitrite by one group of nitrate-reducing microorganisms. Such a production of nitrite will be hidden under the nitrate-limited conditions of most mangrove forest soils.

Phylogenetically diverse anammox bacteria have been detected in most of anoxic natural and engineered ecosystems and thus regarded as key players in the global nitrogen cycle. However, ecological niche differentiation of anammox bacteria remains unresolved despite its ecological and practical importance. In this study, the microbial competitions for a common substrate (nitrite) among three anammox species (i.e. “Candidatus Brocadia sinica”, “Candidatus Jettenia caeni” and “Candidatus Kuenenia stuttgartiensis”) were systematically investigated in nitrite-limited gel-immobilized column reactors (GICR) and membrane bioreactors (MBRs) under different nitrogen loading rates (NLRs). 16 S rRNA gene-based population dynamics revealed that “Ca. J. caeni” could proliferate only at low NLRs, whereas “Ca. B. sinica” outcompeted other two species at higher NLRs in both types of reactors. Furthermore, FISH analysis revealed that “Ca. J. caeni” was mainly present as spherical microclusters at the inner part (low NO2− environment), whereas “Ca. B. sinica” was present throughout the gel beads and granules. This spatial distribution supports the outcomes of the competition experiments. However, the successful competition of “Ca. J. caeni” at low NLR could not be explained with the Monod model probably due to inaccuracy of kinetic parameters such as half saturation constant (Ks) for nitrite and a difference in the maintenance rate (m). In addition, the growth of “Ca. K. stuttgartiensis” could not be observed in any experimental conditions, suggesting possible unknown factor(s) is missing. Taken together, NLR was one of factors determining ecological niche differentiation of “Ca. B. sinica” and “Ca. J. caeni”.

Following initial development of the Grafenberg ring in the 1920's, IUDs fell into disuse until the late 1950s, when plastic devices inserted using new technology began to gain worldwide acceptance. Further research indicated that copper had a significant antifertility effect which increased with increasing surface area, and several copper IUDs were developed and adapted, including the Copper T 200, the Copper T 220C, and the Copper T 380 A, probably the most effective yet. The Gravigard and Multiload are 2 other copper devices developed according to somewhat different principles. Copper devices are widely used not so much because of their great effectiveness as because of their suitability for nulliparous patients and their ease of insertion, which minimizes risk of uterine perforation. Records of 2584 women using Copper IUDs for 7190 women-years and 956 women using devices without copper for 6059 women-years suggest that the copper devices were associated with greater effectiveness and fewer removals for complications. Research suggests that the advantages of copper IUDs become more significant with increased duration of use. Contraindications to copper devices include allergy to copper and hepatolenticular degeneration. No carcinogenic or teratogenic effect of copper devices has been found, but further studies are needed to rule out other undesirable effects. Significant modifications of copper devices in recent years have been developed to increase their effectiveness, prolong their duration of usefulness, facilitate insertion and permit insertion during abortion or delivery. The upper limit of the surface area of copper associated with increased effectiveness appears to be between 200-300 sq mm, and at some point increases in copper exposure may provoke expulsion of the IUD. The duration of fertility inhibition of copper IUDs is usually estimated at 2-3 years, but recent research indicates that it may be 6-8 years, and some devices may retain copper surface

Striped catfish (Pangasianodon hypophthalmus) are farmed intensively at high stocking densities in Vietnam where they are likely to encounter environmental hypercapnia as well as occasional high levels of aquatic nitrite. Nitrite competes with Cl(-) for uptake at the branchial HCO3(-)/Cl(-) exchanger, causing a drastic reduction in the blood oxygen carrying capacity through the formation of methaemoglobin and nitrosylhaemoglobin. Environmental hypercapnia induces a respiratory acidosis where the branchial HCO3(-)/Cl(-) exchange activity is reduced in order to retain HCO3(-) for pH recovery, which should lead to a reduced nitrite uptake. To assess the effect of hypercapnia on nitrite uptake, fish were cannulated in the dorsal aorta, allowing repeated blood sampling for measurements of haemoglobin derivatives, plasma ions and acid-base status during exposure to 0.9mM nitrite alone and in combination with acute and 48h acclimated hypercapnia over a period of 72h. Nitrite uptake was initially reduced during the hypercapnia-induced acidosis, but after pH recovery the situation was reversed, resulting in higher plasma nitrite concentrations and lower functional haemoglobin levels that eventually caused mortality. This suggests that branchial HCO3(-)/Cl(-) exchange activity is reduced only during the initial acid-base compensation, but subsequently increases with the greater availability of internal HCO3(-) counter-ions as pH is compensated. The data further suggest that branchial Na(+)/H(+) exchange plays a significant role in the initial phase of acid-base compensation. Overall, longer term environmental hypercapnia does not protect against nitrite uptake in P. hypophthalmus, but instead enhances it. In addition, we observed a significant size effect in nitrite accumulation, where large fish attained plasma [nitrite] above the ambient concentration, while small fish did not. Small P. hypophthalmus instead had significantly higher plasma [nitrate], and haemoglobin

Full Text Available Sodium nitrite, a common food additive, exists widely not only in the environment but also in our body. Excessive nitrite causes toxicological effects on human health; however, whether it affects vertebrate heart valve development remains unknown. In vertebrates, developmental defects of cardiac valves usually lead to congenital heart disease. To understand the toxic effects of nitrite on valvulogenesis, we exposed zebrafish embryos with different concentrations of sodium nitrite. Our results showed that sodium nitrite caused developmental defects of zebrafish heart dose dependently. It affected zebrafish heart development starting from 36 hpf (hour post fertilization when heart initiates looping process. Comprehensive analysis on the embryos at 24 hpf and 48 hpf showed that excessive nitrite did not affect blood circulation, vascular network, myocardium and endocardium development. But development of endocardial cells in atrioventricular canal (AVC of the embryos at 48 hpf was disrupted by too much nitrite, leading to defective formation of primitive valve leaflets at 76 hpf. Consistently, excessive nitrite diminished expressions of valve progenitor markers including bmp4, has2, vcana and notch1b at 48 hpf. Furthermore, 3', 5'-cyclic guanosine monophosphate (cGMP, downstream of nitric oxide (NO signaling, was increased its level significantly in the embryos exposed with excessive nitrite and microinjection of soluble guanylate cyclase inhibitor ODQ (1H-[1], [2], [4]Oxadiazolo[4,3-a] quinoxalin-1-one, an antagonist of NO signaling, into nitrite-exposed embryos could partly rescue the cardiac valve malformation. Taken together, our results show that excessive nitrite affects early valve leaflet formation by producing too much NO signaling.

The periplasmic nitrate reductase from Cupriavidus necator (also known as Ralstonia eutropha) is a heterodimer that is able to reduce nitrate to nitrite. It comprises a 91 kDa catalytic subunit (NapA) and a 17 kDa subunit (NapB) that is involved in electron transfer. The larger subunit contains a molybdenum active site with a bis-molybdopterin guanine dinucleotide cofactor as well as one [4Fe–4S] cluster, while the small subunit is a di-haem c-type cytochrome. Crystals of the oxidized form of this enzyme were obtained using polyethylene glycol 3350 as precipitant. A single crystal grown at the High Throughput Crystallization Laboratory of the EMBL in Grenoble diffracted to beyond 1.5 Å at the ESRF (ID14-1), which is the highest resolution reported to date for a nitrate reductase. The unit-cell parameters are a = 142.2, b = 82.4, c = 96.8 Å, β = 100.7°, space group C2, and one heterodimer is present per asymmetric unit. PMID:17554176

A new optosensor for visual quantitation of nitrite (NO2(-)) ion has been fabricated by physically immobilizing Safranine O (SO) reagent onto a self-adhesive poly(n-butyl acrylate) [poly(nBA)] microspheres matrix, which was synthesized via facile microemulsion UV lithography technique. Evaluation and optimization of the optical NO2(-) ion sensor was performed with a fiber optic reflectance spectrophotometer. Scanning electron micrograph showed well-shaped and smooth spherical morphology of the poly(nBA) microspheres with a narrow particles size distribution from 0.6 μm up to 1.8 μm. The uniform size distribution of the acrylic microspheres promoted homogeneity of the immobilized SO reagent molecules on the microspheres' surfaces, thereby enhanced the sensing response reproducibility (<5% RSD) with a linear range obtained from 10 to 100 ppm NO2(-) ion. The micro-sized acrylic immobilization matrix demonstrated no significant barrier for diffusion of reactant and product, and served as a good solid state ion transport medium for reflectometric nitrite determination in food samples.

Full Text Available In the present study the nitrogen transport mechanism in plasma nitrited CoCr alloy at moderate temperature ( 400ºC is explained by non-Fickian diffusion model. This mechanism is considered by stress induced diffusion model. The model involves diffusion of nitrogen induced by internal stresses created during nitriding process. The model considers the diffusion of nitrogen in the presence of internal stresses gradient induced by penetrating nitrogen as the next driving force of diffusion after concentration gradient. This model is commonly used for analysis of stainless steel nitriding, however, in this work it is shown that the same nitrogen penetration mechanism takes place in CoCr alloy. For mathematical description of stress induced diffusion process the equation of baro-diffusion is used which involves concentration dependant baro-diffusion concentration. For calculation of stress gradient it is assumed that stress depth profile linearly relates with nitrogen concentration depth profile. The fitting is done using experimental curves of nitrogen depth profiles for medical grade CoCr alloy (ISO 5831-12 nitrited at 400 ºC temperature. The experimental curves are taken from literature. The nitriding duration was 2h, 6h, 20h. Calculated nitrogen depth profiles in CoCr alloy are in good agreement with experimental nitrogen depth profiles. The diffusion coefficient D is found from fitting of experimental data.DOI: http://dx.doi.org/10.5755/j01.ms.21.1.5711

Reaction of 3-5 carbon sugars, glycolaldehyde, and alpha-ketoaldehydes with nitrite under mild anaerobic aqueous conditions yielded ammonia, an essential substrate for the synthesis of nitrogen-containing molecules during abiogenesis. Under the same conditions, ammonia synthesis was not driven by formaldehyde, glyoxylate, 2-deoxyribose, and glucose, a result indicating that the reduction process requires an organic reductant containing either an accessible alpha-hydroxycarbonyl group or an alpha-dicarbonyl group. Small amounts of aqueous Fe(+3) catalyzed the sugar-driven synthesis of ammonia. The glyceraldehyde concentration dependence of ammonia synthesis, and control studies of ammonia's reaction with glyceraldehyde, indicated that ammonia formation is accompanied by incorporation of part of the synthesized ammonia into sugar-derived organic products. The ability of sugars to drive the synthesis of ammonia is considered important to abiogenesis because it provides a way to generate photochemically unstable ammonia at sites of sugar-based origin-of-life processes from nitrite, a plausible prebiotic nitrogen species.

This study introduces a novel approach that uses the interaction of pollutants with added nitrite to produce diazonium salts, which cause in situ self-assembly of the pollutants on carbon electrodes, to improve their 2-fluoroaniline (2-FA) defluorination and removal performance. The 2-FA degradation performance, electrode properties, electrochemical properties and degradation pathway were investigated. The reactor containing NO2(-) achieved a 2-FA removal efficiency of 90.1% and a defluorination efficiency of 38% within 48 h, 1.4 and 2.3 times higher than the corresponding results achieved without NO2(-), respectively. The residual NO2(-) was less than 0.5mg/L in the reactor containing added NO2(-), which would not cause serious secondary pollution. Scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) results proved that the carbon anode surface was successfully modified with benzene polymer, and electrochemical tests confirmed that the electrochemical activity of the modified anode was enhanced significantly. The C-F bond was weakened by the effect of the positive charge of the benzenediazonium groups, and the high electrochemical activity of the carbon anode enhanced the electrochemical performance of the system to accelerate defluorination. Thus, the present electrical method involving nitrite nitrogen is very promising for the treatment of wastewater containing fluoroaniline compounds.

This method provides a procedure for determining nitrate and nitrite concentrations in estuarine and coastal waters. Nitrate is reduced to nitrite by cadmium,1-3 and the resulting nitrite determined by formation of an azo dye.4-6

Two patients with dihydropteridine reductase (DHPR) deficiency, in one case due to the absence of any enzyme protein (DHPR- cross reactive material (CRM)-) and in the other case due to the production of a mutant type devoid of catalytic activity (DHPR- CRM+) were examined. This latter form of malignant phenylketonuria, whose relative frequency seems to be higher in the Italian population, possibly has a worse prognosis. The earlier onset and the greater severity of clinical symptoms are associated with a more pronounced hydroxylation defect, as shown by higher degree of neonatal hyperphenylalaninaemia, unresponsiveness to an oral tetrahydrobiopterin load, lower concentrations of neurotransmitter metabolites, and reduced tyrosine production after an oral phenylalanine load.

Renal injury induced by brain death is characterized by ischemia and inflammation, and limiting it is a therapeutic goal that could improve outcomes in kidney transplantation. Brain death resulted in decreased circulating nitrite levels and increased infiltrating inflammatory cell infiltration into the kidney. Since nitrite stimulates nitric oxide signaling in ischemic tissues, we tested whether nitrite therapy was beneficial in a rat model of brain death followed by kidney transplantation. Nitrite, administered over 2 h of brain death, blunted the increased inflammation without affecting brain death-induced alterations in hemodynamics. Kidneys were transplanted after 2 h of brain death and renal function followed over 7 days. Allografts collected from nitrite-treated brain-dead rats showed significant improvement in function over the first 2 to 4 days after transplantation compared with untreated brain-dead animals. Gene microarray analysis after 2 h of brain death without or with nitrite therapy showed that the latter significantly altered the expression of about 400 genes. Ingenuity Pathway Analysis indicated that multiple signaling pathways were affected by nitrite, including those related to hypoxia, transcription, and genes related to humoral immune responses. Thus, nitrite therapy attenuates brain death-induced renal injury by regulating responses to ischemia and inflammation, ultimately leading to better post-transplant kidney function.

In this paper, thionine was electro-polymerized onto the surface of carbon nanotube (CNT)-modified glassy carbon (GC) to fabricate the polythionine (PTH)/CNT/GC electrode. It was found that the electro-reduction current of nitrite was enhanced greatly at the PTH/CNT/GC electrode. It may be demonstrated that PTH was used as a mediator for electrocatalytic reduction of nitrite, and CNTs as an excellent nanomaterial can improve the electron transfer between the electrode and nitrite. Therefore, based on the synergic effect of PTH and CNTs, the PTH/CNT/GC electrode was employed to detect nitrite, and the high sensitivity of 5.81 {mu}A mM{sup -1}, and the detection limit of 1.4 Multiplication-Sign 10{sup -6} M were obtained. Besides, the modified electrode showed an inherent stability, fast response time, and good anti-interference ability. These suggested that the PTH/CNT/GC electrode was favorable and reliable for the detection of nitrite. - Highlights: Black-Right-Pointing-Pointer Polythionine (PTH) was used as a mediator for electrocatalytic reduction of nitrite. Black-Right-Pointing-Pointer Carbon nanotubes (CNTs) improve electron transfer between the electrode and nitrite. Black-Right-Pointing-Pointer The PTH/CNT/glassy carbon electrode showed excellent nitrite detection performance.

Several food additives are added in food for their preservation to maintain the freshness of food (antioxidants) or to slow down or stop the growth of microorganisms (preservative agents). Nitrites and nitrates are used as preservative agents in meat. Nitrites give a smoked taste, a pinkish color in the meat and protect the consumers against the risk of bacterial deterioration. Their addition is however very limited as, in high dose, it can have risks on human health and the environment. Nitrites may also combine with secondary or tertiary amines to form N-nitroso derivatives. Certain N-nitroso compounds have been shown to produce cancers in a wide range of laboratory animals. Thus, alternatives of nitrates and nitrites are the object of numerous research studies. Alternatives, such as the addition of vitamins, fruits, chemicals products, natural products containing nitrite or spices, which have similar properties of nitrites, are in evaluation. In fact, spices are considered to have several organoleptic and anti-microbial properties which would be interesting to study. Several spices and combinations of spices are being progressively evaluated. This review discusses the sources of nitrites and nitrates, their use as additives in food products, their physicochemical properties, their negatives effects and the use of alternatives of nitrites and nitrates in preserving meat products.

Nitrite intake from locally processed meat in Khartoum, Sudan was estimated and compared to established safety levels. For this purpose, 90 locally processed meat samples were collected randomly from retail outlets and analysed for nitrite levels according to the British standard 1992 protocol and 3

Sulfur-limestone was used in the autotrophic denitrification process to remove the nitrate and nitrite in a lab scale upflow biofilter.Synthetic water with four levels of nitrate and nitrite concentrations of 10,40,70 and 100 mg N/L was tested.When treating the low concentration of nitrate- or nitrite-contaminated water (10,40 mg N/L),a high removal rate of about 90% was achieved at the hydraulic retention time (HRT) of 3 hr and temperature of 20-25℃.At the same HRT,50% of the nitrate or nitrite could be removed even at the low temperature of 5-10℃.For the higher concentration nitrate and nitrite (70,100 mg N/L),longer HRT was required.The batch test indicated that influent concentration,HRT and temperature are important factors affecting the denitrification efficiency.Molecular analysis implied that nitrate and nitrite were denitrified into nitrogen by the same microorganisms.The sequential two-stepreactions from nitrate to nitrite and from nitrite to the next-step product might have taken place in the same cell during the autotrophic denitrification process.

The mechanism of nitrite hydrogenation over a Pd/Al2O3 catalyst layer deposited on a ZnSe internal reflection element was investigated in water using attenuated total reflection infrared spectroscopy. Nitrite hydrogenates to NO(ads), NH2(ads), and NH+4 on the palladium surface. Hydrogenation of

The tolerance and effects of nitrite on ion balance and haematology were investigated in the striped snakehead, Channa striata Bloch 1793, which is an air-breathing fish with reduced gills of importance for aquaculture in South East Asia. C. striata was nitrite tolerant with a 96 h LC50 of 4.7 m...

Chloride induced-corrosion of steel bars in concrete can make cracks and exfoliation in near-surface regions in reinforced concrete structures. In this paper, we described the basic concept and practice of steel bars corrosion protection method by the combination of galvanic anode (zinc wire) and the penetration of nitrite ions from mortar layers containing a large amount of lithium nitrite.

substantial amounts. Studies were made of the reactions of ethoxyl radicals (produced by the decomposition of diethyl peroxide at 181 deg C) with ethyl nitrite...and NO. The reaction rate between the ethoxyl radicals and NO was much greater than that between the radicals the nitrite.

Nitrite may be a source for nitric oxide (*NO), particularly in highly acidic environments, such as the stomach. Diet products contribute also with reductants that dramatically increase the production of *NO from nitrite. Red wine has been attributed health promoting properties largely on basis of the reductive antioxidant properties of its polyphenolic fraction. We show in vitro that wine, wine anthocyanin fraction and wine catechol (caffeic acid) dose- and pH-dependently promote the formation of *NO when mixed with nitrite, as measured electrochemically. The production of *NO promoted by wine from nitrite was substantiated in vivo in healthy volunteers by measuring *NO in the air expelled from the stomach, following consumption of wine, as measured by chemiluminescence. Mechanistically, the reaction involves the univalent reduction of nitrite, as suggested by the formation of *NO and by the appearance of EPR spectra assigned to wine phenolic radicals. Ascorbic and caffeic acids cooperate in the reduction of nitrite to *NO. Moreover, reduction of nitrite is critically dependent on the phenolic structure and nitro-derivatives of phenols are also formed, as suggested by caffeic acid UV spectral modifications. The reduction of nitrite may reveal previously unrecognized physiologic effects of red wine in connection with *NO bioactivity.

A fully automated procedure based on Sequential Injection Analysis (SIA) methodology for simultaneous monitoring of nitrate and nitrite in surface water samples is described. Nitrite was determined directly using the Griess diazo-coupling reaction and the formed azo dye was measured at 540 nm in the flow cell of the fibre-optic spectrophotometer. Nitrate zone was passed through a reducing mini-column containing copperised-cadmium. After the reduction of nitrate into nitrite the sample was aspirated by flow reversal to the holding coil, treated with the reagent and finally passed through the flow cell. The calibration curve was linear over the range 0.05-1.00 mg N l(-1) of nitrite and 0.50-50.00 mg N l(-1) of nitrate; correlation coefficients were 0.9993 and 0.9988 for nitrite and nitrate, respectively. Detection limits were 0.015 and 0.10 mg N l(-1) for nitrite and nitrate, respectively. The relative standard deviation (RSD) values (n = 3) were 1.10% and 1.32% for nitrite and nitrate, respectively. The total time of one measuring cycle was 250 s, thus the sample throughput was about 14 h(-1). Nitrate and nitrite were determined in the real samples of surface water, and the results have been compared with those obtained by two other flow methods; flow injection analysis based on the same reactions and isotachophoretic determination used in a routine environmental control laboratory.

was converted to nitrite, with the average total net N2O production of 2.1 ± 0.7% of the ammonium oxidized. Operation with intermittent feeding appears an effective optimization approach to mitigate N2O emissions from nitritating systems. Net N2O production rates transiently increased with a rise in pH after...

Full Text Available Nitrite (NO2– is a substrate for both oxidative and reductive microbial metabolism. NO2– accumulates at the base of the euphotic zone in oxygenated, stratified open ocean water columns, forming a feature known as the primary nitrite maximum (PNM. Potential pathways of NO2– production include the oxidation of ammonia (NH3 by ammonia-oxidizing bacteria or archaea and assimilatory nitrate (NO3– reduction by phytoplankton or heterotrophic bacteria. Measurements of NH3 oxidation and NO3– reduction to NO2– were conducted at two stations in the central California Current in the eastern North Pacific to determine the relative contributions of these processes to NO2– production in the PNM. Sensitive (−1, high-resolution measurements of [NH4+] and [NO2–] indicated a persistent NH4+ maximum overlying the PNM at every station, with concentrations as high as 1.5 μmol L−1. Within and just below the PNM, NH3 oxidation was the dominant NO2– producing process with rates of NH3 oxidation of up to 50 nmol L−1 d−1, coinciding with high abundances of ammonia-oxidizing archaea. Though little NO2– production from NO3– was detected, potentially nitrate-reducing phytoplankton (photosynthetic picoeukaryotes, Synechococcus, and Prochlorococcus were present at the depth of the PNM. Rates of NO2– production from NO3– were highest within the upper mixed layer (4.6 nmol L−1 d−1 but were either below detection limits or 10 times lower than NH3 oxidation rates around the PNM. One-dimensional modeling of water column NO2– profiles supported direct rate measurements of a net biological sink for NO2– just below the PNM. Residence time estimates of NO2– within the PNM were similar at the mesotrophic and oligotrophic stations and ranged from 150–205 d. Our results suggest the PNM is a dynamic, rather than relict, feature with a source term dominated by ammonia oxidation.

We investigated the possible use of atmospheric pressure plasma-treated water (PTW) as a nitrite source in curing process. Emulsion-type sausages were manufactured with PTW, celery powder containing nitrite, and synthetic sodium nitrite at a concentration of nitrite ion 70mgkg(-1). In terms of sausage quality, there were no noticeable effects of PTW on the total aerobic bacterial counts, color, and peroxide values of sausages compared with those of celery powder and sodium nitrite throughout 28days of storage at 4°C. Sausage with added PTW had lower concentrations of residual nitrite compared to those of added celery powder and sodium nitrite during the storage period (Psausages were not different, whereas the sausage with added celery powder received the lowest scores in taste and acceptability. From the results, it is concluded that PTW can be used as a nitrite source equivalent to a natural curing agent.

The effects of sodium nitrite and ascorbate on lipid and protein oxidation were studied during the ripening process of dry fermented sausages. Samples were taken at day 0, 2, 8, 14, 21 and 28 of ripening to assess lipid (malondialdehyde) and protein (carbonyls and sulfhydryl groups) oxidation. Sodium ascorbate and nitrite were separately able to reduce the formation of malondialdehyde. Their combined addition resulted in higher amounts of carbonyl compounds compared to their separate addition or the treatment without any of both compounds. Moreover, sodium nitrite limited the formation of γ-glutamic semialdehyde whereas sodium ascorbate showed a pro-oxidant effect. A loss of thiol groups was observed during ripening, which was not affected by the use of sodium ascorbate nor sodium nitrite. In conclusion, sodium nitrite and ascorbate affected protein and lipid oxidation in different manners. The possible pro-oxidant effect of their combined addition on carbonyl formation might influence the technological and sensory properties of these products.

The amount of piggery wastewater as domestic livestock is increasing.The volume of piggery wastewater produced is less than the volume of other wastewaters,but piggery wastewater has a heavy impact on wastewater streams due to an extremely high concentration of nitrogen and COD.In this study,laboratory reactors were operated using piggery wastewater and the effluent of anaerobic digester from piggery wastewater plants.The purpose of this study was to induce the nitritation process,which is an economically advantageous nitrogen removal method that converts ammonium nitrogen into nitrite.The results showed that the effluent of anaerobic digester from piggery wastewater was more efficient than raw piggery wastewater in terms of inducing nitritation.It can be deduced that nitritation is largely affected by an organic fraction of piggery wastewater.It can also be concluded that a small amount of biodegradable organic matter in piggery wastewater is efficient in inducing nitritation.

A major percentage of fixed nitrogen (N) loss in the oceans occurs within nitrite-rich oxygen minimum zones (OMZs) via denitrification and anammox. It remains unclear to what extent ammonium and nitrite oxidation co-occur, either supplying or competing for substrates involved in nitrogen loss in the OMZ core. Assessment of the oxygen (O2) sensitivity of these processes down to the O2 concentrations present in the OMZ core (Michaelis-Menten model, indicating a high-affinity component with a Km of just a few nanomolar. As the communities of ammonium and nitrite oxidizers were similar to other OMZs, these kinetics should apply across OMZ systems. The high O2 affinities imply that ammonium and nitrite oxidation can occur within the OMZ core whenever O2 is supplied, for example, by episodic intrusions. These processes therefore compete with anammox and denitrification for ammonium and nitrite, thereby exerting an important control over nitrogen loss.

Copper is an essential trace element for many important cellular functions. However, excess of copper can impair cellular functions by copper-induced oxidative stress. In brain, astrocytes are considered to play a prominent role in the copper homeostasis. In this short review we summarise the current knowledge on the molecular mechanisms which are involved in the handling of copper by astrocytes. Cultured astrocytes efficiently take up copper ions predominantly by the copper transporter Ctr1 and the divalent metal transporter DMT1. In addition, copper oxide nanoparticles are rapidly accumulated by astrocytes via endocytosis. Cultured astrocytes tolerate moderate increases in intracellular copper contents very well. However, if a given threshold of cellular copper content is exceeded after exposure to copper, accelerated production of reactive oxygen species and compromised cell viability are observed. Upon exposure to sub-toxic concentrations of copper ions or copper oxide nanoparticles, astrocytes increase their copper storage capacity by upregulating the cellular contents of glutathione and metallothioneins. In addition, cultured astrocytes have the capacity to export copper ions which is likely to involve the copper ATPase 7A. The ability of astrocytes to efficiently accumulate, store and export copper ions suggests that astrocytes have a key role in the distribution of copper in brain. Impairment of this astrocytic function may be involved in diseases which are connected with disturbances in brain copper metabolism.

Background Salivary nitrate is positively correlated with plasma nitrate and its level is 9 times the plasma level after nitrate loading. Nitrate in saliva is known to be reduced to nitrite by oral bacteria. Nitrate and nitrite levels in saliva are 3-5 times those in serum in physiological conditions respectively in our previous study. The biological functions of high salivary nitrate and nitrite are still not well understood. The aim of this in vitro study was to investigate the antimicrobial effects of nitrate and nitrite on main oral pathogens under acidic conditions.Methods Six common oral pathogens including Streptococcus mutans NCTC 10449, Lactobacillus acidophilus ATCC 4646, Porphyromonas gingivalis ATCC 33277, Capnocytophaga gingivalis ATCC 33624, Fusobacterium nucleatum ATCC 10953, and Candida albicans ATCC 10231 were cultured in liquid medium. Sodium nitrate or sodium nitrite was added to the medium to final concentrations of 0, 0.5, 1, 2, and 10 mmol/L. All of the microorganisms were incubated for 24 to 48 hours. The optical densities (OD) of cell suspensions were determined and the cultures were transferred to solid nutrient broth medium to observe the minimum inhibitory concentration and minimum bactericidal/fungicidal concentration for the six tested pathogens.Results Nitrite at concentrations of 0.5 to 10 mmol/L had an inhibitory effect on all tested organisms at low pH values. The antimicrobial effect of nitrite increased with the acidity of the medium. Streptococcus mutans NCTC 10449 was highly sensitive to nitrite at low pH values. Lactobacillus acidophilus ATCC 4646 and Candida albicans ATCC 10231 were relatively resistant to acidified nitrite. Nitrate at the given concentrations and under acidic conditions had no inhibitory effect on the growth of any of the tested pathogens.Conclusion Nitrite, at a concentration equal to that in human saliva, is both cytocidal and cytostatic to six principal oral pathogens in vitro, whereas nitrate at a similar

Monitoring of nitrite is essential for an immediate response and prevention of irreversible failure of decentralized biological urine nitrification reactors. Although a few sensors are available for nitrite measurement, none of them are suitable for applications in which both nitrite and nitrate are present in very high concentrations. Such is the case in collected source-separated urine, stabilized by nitrification for long-term storage. Ultraviolet (UV) spectrophotometry in combination with chemometrics is a promising option for monitoring of nitrite. In this study, an immersible in situ UV sensor is investigated for the first time so to establish a relationship between UV absorbance spectra and nitrite concentrations in nitrified urine. The study focuses on the effects of suspended particles and saturation on the absorbance spectra and the chemometric model performance. Detailed analysis indicates that suspended particles in nitrified urine have a negligible effect on nitrite estimation, concluding that sample filtration is not necessary as pretreatment. In contrast, saturation due to very high concentrations affects the model performance severely, suggesting dilution as an essential sample preparation step. However, this can also be mitigated by simple removal of the saturated, lower end of the UV absorbance spectra, and extraction of information from the secondary, weaker nitrite absorbance peak. This approach allows for estimation of nitrite with a simple chemometric model and without sample dilution. These results are promising for a practical application of the UV sensor as an in situ nitrite measurement in a urine nitrification reactor given the exceptional quality of the nitrite estimates in comparison to previous studies.

Full Text Available Nitrites are potential carcinogens. Therefore, limiting nitrites in food is critically important for food safety. The nitrite degradation capacity of Lactobacillus casei subsp. rhamnosus LCR 6013 was investigated in pickle fermentation. After LCR 6013 fermentation for 120 h at 37°C, the nitrite concentration in the fermentation system was significantly lower than that in the control sample without the LCR 6013 strain. The effects of NaCl and Vc on nitrite degradation by LCR 6013 in the De Man, Rogosa and Sharpe (MRS medium were also investigated. The highest nitrite degradations, 9.29 mg/L and 9.89 mg/L, were observed when NaCl and Vc concentrations were 0.75% and 0.02%, respectively in the MRS medium, which was significantly higher than the control group (p ≤ 0.01. Electron capture/gas chromatography and indophenol blue staining were used to study the nitrite degradation pathway of LCR 6013. The nitrite degradation products contained N2O, but no NH4(+. The LCR 6013 strain completely degraded all NaNO2 (50.00 mg/L after 16 h of fermentation. The enzyme activity of NiR in the periplasmic space was 2.5 times of that in the cytoplasm. Our results demonstrated that L. casei subsp. rhamnosus LCR 6013 can effectively degrade nitrites in both the pickle fermentation system and in MRS medium by NiR. Nitrites are degraded by the LCR 6013 strain, likely via the nitrate respiration pathway (NO2(->NO->N2O->N2, rather than the aammonium formation pathway (dissimilatory nitrate reduction to ammonium, DNRA, because the degradation products contain N2O, but not NH4(+.

The potential activity of nitrate reductase was determined in uni-algal cultures in the laboratory and in natural marine phytoplankton assemblages. In the laboratory bioassays, distinct differences in nitrate reductase activity were observed in iron replete versus depleted cultures for Emiliania hux

Hereditary forms of copper toxicosis exist in man and dogs. In man, Wilson’s disease is the best studied disorder of copper overload, resulting from mutations in the gene coding for the copper transporter ATP7B. Forms of copper toxicosis for which no causal gene is known yet are recognized as well, often in young children. Although advances have been made in unraveling the genetic background of disorders of copper metabolism in man, many questions regarding disease mechanisms and copper homeo...

Iron and copper are essential trace metals, actively absorbed from the proximal gut in a regulated fashion. Depletion of either metal can lead to anemia. In the gut, copper deficiency can affect iron absorption through modulating the activity of hephaestin - a multi-copper oxidase required for optimal iron export from enterocytes. How systemic copper status regulates iron absorption is unknown. Mice were subjected to a nutritional copper deficiency-induced anemia regime from birth and injected with copper sulphate intraperitoneally to correct the anemia. Copper deficiency resulted in anemia, increased duodenal hypoxia and Hypoxia inducible factor 2α (HIF-2α) levels, a regulator of iron absorption. HIF-2α upregulation in copper deficiency appeared to be independent of duodenal iron or copper levels and correlated with the expression of iron transporters (Ferroportin - Fpn, Divalent Metal transporter - Dmt1) and ferric reductase - Dcytb. Alleviation of copper-dependent anemia with intraperitoneal copper injection resulted in down regulation of HIF-2α-regulated iron absorption genes in the gut. Our work identifies HIF-2α as an important regulator of iron transport machinery in copper deficiency.

Nitrate and nitrite are authorised as preservatives in certain food products, such as salami, ham (nitrite) and cheese (nitrate). They prevent food spoilage and protect the consumer against food-borne pathogens. Next to that, nitrate and nitrite play a role in food colour retention and contribute to

... International Trade Administration Sodium Nitrite From the People's Republic of China: Final Results of the... countervailing duty (``CVD'') order on sodium nitrite from the People's Republic of China (``PRC'') would be... sodium nitrite from the PRC, pursuant to section 751(c) of the Tariff Act of 1930, as amended (``the...

... International Trade Administration Sodium Nitrite From Germany and the People's Republic of China: Final Results... reviews of the antidumping duty (``AD'') orders on sodium nitrite from Germany and the People's Republic... nitrite from Germany and the PRC.\\1\\ On July 1, 2013, the Department published the notice of initiation...

... COMMISSION Sodium Nitrite From China And Germany; Scheduling of Expedited Five-Year Reviews Concerning the... on Sodium Nitrite From Germany AGENCY: United States International Trade Commission. ACTION: Notice... duty order on sodium nitrite from Germany would be likely to lead to continuation or recurrence...

Hypochlorous acid (HOCl) is elevated in many inflammatory diseases and causes the accumulation of free iron. Through the Fenton reaction, free iron has the ability to generate free radicals and subsequently is toxic. Recent studies have demonstrated that HOCl participates in heme destruction of hemoglobin (Hb) and free iron release. In this study, it was showed that nitrite (NO2(-)) could prevent HOCl-mediated Hb heme destruction and free iron release. Also, NO2(-) prevented HOCl-mediated loss of Hb peroxidase activity. After the NO2(-)/HOCl treatment, Tyr 42 in α-chain was found to be nitrated in Hb, attenuating the electron transferring abilities of phenolic compounds. The protective effects of NO2(-) on HOCl-induced heme destruction were attributed to its reduction of ferryl Hb and/or direct scavenging of HOCl. Therefore, NO2(-) could show protective effects in some inflammatory diseases by preventing HOCl-mediated heme destruction of hemoproteins and free iron release.

The antifertility effect of copper-bearing IUDs is based on continuous release of copper, which is a result of the reaction between the metal and the uterine secretions. Released cupric ions collect in the endometrium and in the uterine fluid but significant accumulation has not been found in the bloodstream or elsewhere. Following Laker's suggestion that hair be used for monitoring essential trace elements, e.g., copper, we checked the copper content of the hair of women wearing copper-bearing IUDs. Samples of untreated pubic hair removed by clipping before diagnostic curettage were obtained from 10 young (24-34 years old), white caucasian females who until then had been wearing an MLCu250 IUD for more than 1 year. Pubes from 10 comparable (sex, age, race) subjects who had never used a Cu-containing device served as controls. The unwashed material was submitted to the toxicology laboratory, where the copper content was assessed by flameless atomic absorption, a technique whose lower limit of measurement lies at a concentration of 0.05 mcg Cu/ml fluid (50 ppb). Hair samples were washed to remove extraneous traces of metal according to the prescriptions of the International Atomic Energy Agency, weighed, and mineralized, after which a small volume (10 mcl) of the diluted fluid was fed into the graphite furnace. Each sample (75-150 mg) was analyzed 4 times, both before and after washing. Since the cleaning procedure reduces the weight of the sample (mainly by the removal of fat, dust, etc.) this explains why the percentage copper content of washed hair is higher than that of unwashed hair belonging to the same subject. The results indicate that there was no significant difference (Mann-Whitney U test) between the mean copper levels of both unwashed and washed pubes from women who were using or had never used an MLCu250 IUD. We therefore conclude that the use of this copper-containing device is not associated with significant accumulation of copper in (pubic) hair.

Highlights: • A method for improving defluorination performance by in situ self-assembly of pollutants was developed. • The mechanisms of 2-FA modification and defluorination are discussed. • Positively-charged diazonium salt is used to weaken the C–F bond. - Abstract: This study introduces a novel approach that uses the interaction of pollutants with added nitrite to produce diazonium salts, which cause in situ self-assembly of the pollutants on carbon electrodes, to improve their 2-fluoroaniline (2-FA) defluorination and removal performance. The 2-FA degradation performance, electrode properties, electrochemical properties and degradation pathway were investigated. The reactor containing NO{sub 2}{sup −} achieved a 2-FA removal efficiency of 90.1% and a defluorination efficiency of 38% within 48 h, 1.4 and 2.3 times higher than the corresponding results achieved without NO{sub 2}{sup −}, respectively. The residual NO{sub 2}{sup −} was less than 0.5 mg/L in the reactor containing added NO{sub 2}{sup −}, which would not cause serious secondary pollution. Scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) results proved that the carbon anode surface was successfully modified with benzene polymer, and electrochemical tests confirmed that the electrochemical activity of the modified anode was enhanced significantly. The C–F bond was weakened by the effect of the positive charge of the benzenediazonium groups, and the high electrochemical activity of the carbon anode enhanced the electrochemical performance of the system to accelerate defluorination. Thus, the present electrical method involving nitrite nitrogen is very promising for the treatment of wastewater containing fluoroaniline compounds.

1. Market Consumption The ’China Factor’ and Copper Price Fluctuation We all know China is an enormous consumer of copper,but the exact levels of consumption and where the copper has gone remains a mystery.

Copper nanoparticles have been the focus of intensive study due to their potential applications in diverse fields including biomedicine, electronics, and optics. Copper-based nanostructured materials have been used in conductive films, lubrification, nanofluids, catalysis, and also as potent antimicrobial agent. The biogenic synthesis of metallic nanostructured nanoparticles is considered to be a green and eco-friendly technology since neither harmful chemicals nor high temperatures are involved in the process. The present review discusses the synthesis of copper nanostructured nanoparticles by bacteria, fungi, and plant extracts, showing that biogenic synthesis is an economically feasible, simple and non-polluting process. Applications for biogenic copper nanoparticles are also discussed.

Water column nitrification is a key process in the nitrogen cycle as it links reduced and oxidized forms of nitrogen and also provides the substrate (nitrate) needed for reactive nitrogen removal by denitrification. We measured potential water column ammonium and nitrite oxidation rates at four sites along an estuary to continental shelf gradient over two summers. In most cases, nitrite oxidation rates outpaced ammonium oxidation rates. Overall, ammonium and nitrite oxidation rates were higher outside of the estuary, and this trend was primarily driven by higher oxidation rates in deeper waters. Additionally, both ammonium and nitrite oxidation rates were impacted by different in situ variables. Ammonium oxidation rates throughout the water column as a whole were most positively correlated to depth and salinity and negatively correlated to dissolved oxygen, light, and temperature. In contrast, nitrite oxidation rates throughout the water column were negatively correlated with temperature, light and pH. Multivariate regression analysis revealed that surface (20 m) rates were regulated by temperature, light, and [H+] (i.e. pH). In addition, surface (oxygen all played a role in predicting deep (>20 m) nitrite oxidation rates. These results support the growing body of evidence that ammonium oxidation and nitrite oxidation are not always coupled, should be measured separately, and are influenced by different environmental conditions.

During 1985～1987, the concentration of nitrate nitrogen was higher in the Laizhou Bay and the Bohai Bay while that of nitrite nitrogen was higher in the Liaodong Bay and the Bohai Bay. The concentration of nitrate nitrogen was highest in winter and lowest in summer while that of nitrite nitrogen was highest in autumn and lowest in spring. The seasonal variation of the concentration of nitrate nitrogen was maximum in the Laizhou Bay and the Bohai Bay while that of the concentration of nitrite nitrogen was maximum in the Liaodong Bay. There was a great difference in the concentration of nitrate nitrogen between the surface and the bottom in autumn and in the concentration of nitrite nitrogen between the surface and the bottom in summer. The main reason for the seasonal variations of the concentration of nitrate nitrogen and nitrite nitrogen was the marine biochemical process. The nitrate nitrogen and nitrite nitrogen in the Bohai Sea basically maintained a quasi-equilibrium state seasonal cycle. The quasi-equilibrium state seasonal cycle of nitrate nitrogen and nitrite nitrogen at the bottom was stable while that at the surface was liable to variations caused by other factors.

Environmental chloride has been shown to inhibit methemoglobin formation in fish, thereby offering a protective effect against nitrite toxicity. Channel catfish (Ictalurus punctatus) were simultaneously exposed to various environmental nitrite and chloride levels (as either CaCl2 or NaCl) in dechlorinated tap water (40 mg/L total hardness, 47 mg/L alkalinity, 4 mg/L chloride, pH = 6.9-7.1, and temperature 21-24°C). Methemoglobin levels in fish simultaneously exposed to 2.5 mg/L nitrite and up to 30 mg/L chloride as either CaCl2 or NaCl were similar but significantly lower than in unprotected fish. Exposure to 10 mg/L nitrite and 60 mg/L chloride resulted in methemoglobin levels similar to those of the controls; most unprotected fish died. Fish exposed to 10 mg/L nitrite had significantly lower methemoglobin levels when protected with 15.0 mg/L chloride as CaCl2 than with NaCl. Fish exposed to nitrite in the presence of 60 mg/L chloride (as either CaCl2 or NaCl) had similar 24-h LC50 values that were significantly elevated above those obtained in the absence of chloride. Calcium had little effect on tolerance to nitrite toxicity in channel catfish in contrast to its large effect reported in steelhead trout (Salmo gairdneri).

Water column nitrification is a key process in the nitrogen cycle as it links reduced and oxidized forms of nitrogen and also provides the substrate (nitrate) needed for reactive nitrogen removal by denitrification. We measured potential water column ammonium and nitrite oxidation rates at four sites along an estuary to continental shelf gradient over two summers. In most cases, nitrite oxidation rates outpaced ammonium oxidation rates. Overall, ammonium and nitrite oxidation rates were higher outside of the estuary, and this trend was primarily driven by higher oxidation rates in deeper waters. Additionally, both ammonium and nitrite oxidation rates were impacted by different in situ variables. Ammonium oxidation rates throughout the water column as a whole were most positively correlated to depth and salinity and negatively correlated to dissolved oxygen and light. In contrast, nitrite oxidation rates throughout the water column were negatively correlated with light and pH. Multivariate regression analysis revealed that while both surface (20 m) ammonium oxidation rates were most strongly predicted by depth and light, surface rates were also regulated by salinity and deep rates by temperature. Surface (20 m) nitrite oxidation rates. These results support the growing body of evidence that ammonium oxidation and nitrite oxidation are not always coupled, should be measured separately, and are influenced by different environmental conditions.

The present study was designed to determine the protective effects of Yucca schidigera (Ys) against oxidative damage induced by acute nitrite intoxication as well as the histopathological evaluation of Ys in rats. The rats were divided into three groups each containing 12 rats: control (C); nitrite intoxication (N); Ys + nitrite intoxication (NY). C and N groups were fed standard rat feed (SRF). The NY group was fed SRF + 100 ppm Ys powder for 4 weeks. Acute nitrite intoxication was induced by subcutaneous (s.c.) administration of sodium nitrite (60 mg/kg) 1 day after the feeding period. Fifty minutes after sodium nitrite administration, blood samples and tissues including lung, liver, and kidney were collected for clinical biochemistry and histopathological investigations. Ys treatment was found to decrease methemoglobin, blood and tissue malondialdehyde, and tissue nitric oxide concentrations, and to increase the glutathione in blood and various tissues. However, plasma nitric oxide, total antioxidant activity, beta-carotene, and vitamin A did not differ between N and NY groups. While the N group rats showed distinct pathology in various tissues (compared with controls), the NY group had similar lung and liver pathology to the control. Only moderate or mild hemorrhage and hyperemia were seen in kidneys of NY group rats. Consequently, the natural compounds found in Ys, such as polyphenols, steroidal saponins, and other phytonutrients, could be used to substantially protect the organism from nitrite-induced oxidative damage and its complications.

Highlights: •Acute effects of waterborne copper were evaluated in the estuarine guppy Poecilia vivipara. •Fishes were acutely exposed to waterborne copper in salt water. •Waterborne copper affects the response of several biochemical and genetic endpoints. •Catalase, reactive oxygen species, antioxidant capacity and lipid peroxidation are responsive to copper exposure. •Copper exposure induces DNA damages in fish erythrocytes. -- Abstract: The responses of a large suite of biochemical and genetic parameters were evaluated in tissues (liver, gills, muscle and erythrocytes) of the estuarine guppy Poecilia vivipara exposed to waterborne copper in salt water (salinity 24 ppt). Activities of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase, and glutathione S-transferase), metallothionein-like protein concentration, reactive oxygen species (ROS) content, antioxidant capacity against peroxyl radicals (ACAP), and lipid peroxidation (LPO) were evaluated in liver, gills, and muscle. Comet assay score and nuclear abnormalities and micronucleated cell frequency were analyzed in peripheral erythrocytes. The responses of these parameters were evaluated in fish exposed (96 h) to environmentally relevant copper concentrations (5, 9 and 20 μg L{sup −1}). In control and copper-exposed fish, no mortality was observed over the experimental period. Almost all biochemical and genetic parameters proved to be affected by waterborne copper exposure. However, the response of catalase activity in liver, ROS, ACAP and LPO in muscle, gills and liver, and DNA damages in erythrocytes clearly showed to be dependent on copper concentration in salt water. Therefore, the use of these parameters could be of relevance in the scope of biomonitoring programs in salt water environments contaminated with copper.

The tolerance and effects of nitrite on ion balance and haematology were investigated in the striped snakehead, Channa striata Bloch 1793, which is an air-breathing fish with reduced gills of importance for aquaculture in South East Asia. C. striata was nitrite tolerant with a 96 h LC50 of 4.7 mM. Effects of sub-lethal exposures to nitrite (0mM, 1.4mM, and 3.0mM) were determined during a 7-day exposure period. Plasma nitrite increased, but the internal concentration remained well below ambient levels. Extracellular nitrate rose by several mM, indicating that a large proportion of the nitrite taken up was converted to nitrate. Nitrite reacted with erythrocyte haemoglobin (Hb) causing methaemoglobin (metHb) to increase to 30% and nitrosylhaemoglobin (HbNO) to increase to 10% of total Hb. Both metHb and HbNO stabilised after 4 days, and functional Hb levels accordingly never fell below 60% of total Hb. Haematocrit and total Hb were unaffected by nitrite. Although the effects of nitrite exposure seemed minor in terms of plasma nitrite and metHb increases, ion balance was strongly affected. In the high exposure group, total osmolality decreased from 320 mOsm to 260 mOsm, and plasma sodium from 150 mM to 120 mM, while plasma chloride fell from 105 mM to 60mM and plasma bicarbonate rose from 12 mM in controls to 20mM in exposed fish. The extreme changes in ion balance in C. striata are different from the response reported in other fish, and further studies are needed to investigate the mechanism behind the observed changes in regulation.

The Eastern Tropical South Pacific is one of the three major oxygen deficient zones (ODZs) in the global ocean and is responsible for approximately one third of marine water column nitrogen loss. It is the best studied of the ODZs and, like the others, features a broad nitrite maximum across the low oxygen layer. How the microbial processes that produce and consume nitrite in anoxic waters interact to sustain this feature is unknown. Here we used 15N-tracer experiments to disentangle five of the biologically mediated processes that control the nitrite pool, including a high-resolution profile of nitrogen loss rates. Nitrate reduction to nitrite likely depended on organic matter fluxes, but the organic matter did not drive detectable rates of denitrification to N2. However, multiple lines of evidence show that denitrification is important in shaping the biogeochemistry of this ODZ. Significant rates of anaerobic nitrite oxidation at the ODZ boundaries were also measured. Iodate was a potential oxidant that could support part of this nitrite consumption pathway. We additionally observed N2 production from labeled cyanate and postulate that anammox bacteria have the ability to harness cyanate as another form of reduced nitrogen rather than relying solely on ammonification of complex organic matter. The balance of the five anaerobic rates measured—anammox, denitrification, nitrate reduction, nitrite oxidation, and dissimilatory nitrite reduction to ammonium—is sufficient to reproduce broadly the observed nitrite and nitrate profiles in a simple one-dimensional model but requires an additional source of reduced nitrogen to the deeper ODZ to avoid ammonium overconsumption.

Solubilities were determined for sodium nitrate, sodium nitrite, and sodium aluminate in synthetic nuclear waste liquor. Solubilities were determined as a function of temperature and solution composition (concentrations of sodium hydroxide, sodium nitrate, sodium nitrite, and sodium aluminate). Temperature had the greatest effect on the solubilities of sodium nitrate and sodium nitrite and a somewhat lesser effect on sodium aluminate solubility. Hydroxide had a great effect on the solubilities of all three salts. Other solution components had minor effects. 2 references, 8 figures, 11 tables.

In certain social circles a variety of volatile nitrites are being used extensively to expand creativity, stimulate music appreciation, promote a sense of abandon in dancing, and intensify sexual experience. Soon after amyl nitrite (the prototype of this group) became a prescription drug, legally produced substitutes appeared and were sold as room odorizers. Analyses of eight of these substitutes showed them to contain isomers of butyl or amyl alcohol and a corresponding nitrite. The authors describe the context of use of these products and their desired and adverse effects.

Full Text Available This article describes the susceptibility of 316L stainless steel to stress corrosion cracking (SCC in a nitrite-containing chloride solution. Slow strain rate testing (SSRT in 30 wt. % MgCl2 solution established SCC susceptibility, as evidenced by post-SSRT fractography. Addition of nitrite to the chloride solution, which is reported to have inhibitive influence on corrosion of stainless steels, was found to increase SCC susceptibility. The susceptibility was also found to increase with nitrite concentration. This behaviour is explained on the basis of the passivation and pitting characteristics of 316L steel in chloride solution.

Copper and Silver Halates is the third in a series of four volumes on inorganic metal halates. This volume presents critical evaluations and compilations for halate solubilities of the Group II metals. The solubility data included in this volume are those for the five compounds, copper chlorate and iodate, and silver chlorate, bromate and iodate.

Copper has been intensively used in industry and agriculture since mid-18(th) century and is currently accumulating in soils. We investigated the diversity of potential active bacteria by 16S rRNA gene transcript amplicon sequencing in a temperate grassland soil subjected to century-long exposure......, suggesting a potential promising role as bioindicators of copper contamination in soils....

Sodium nitrite alone is shown to ameliorate sub-lethal cyanide toxicity in mice when given from ~1 hour before until 20 minutes after the toxic dose as demonstrated by the recovery of righting ability. An optimum dose (12 mg/kg) was determined to significantly relieve cyanide toxicity (5.0 mg/kg) when administered to mice intraperitoneally. Nitrite so administered was shown to rapidly produce NO in the bloodsteam as judged by the dose dependent appearance of EPR signals attributable to nitrosylhemoglobin and methemoglobin. It is argued that antagonism of cyanide inhibition of cytochrome c oxidase by NO is the crucial antidotal activity rather than the methemoglobin-forming action of nitrite. Concomitant addition of sodium thiosulfate to nitrite-treated blood resulted in the detection of sulfidomethemoblobin by EPR spectroscopy. Sulfide is a product of thiosulfate hydrolysis and, like cyanide, is known to be a potent inhibitor of cytochrome c oxidase; the effects of the two inhibitors being essentially additive under standard assay conditions, rather than dominated by either one. The findings afford a plausible explanation for an observed detrimental effect in mice associated with the use of the standard nitrite-thiosulfate combination therapy at sub-lethal levels of cyanide intoxication. PMID:21534623

We report a simple method for the stable dispersion of multi-walled carbon nanotubes (MWNTs) in water by vanillin and controllable surface addition onto carbon fiber microelectrodes (CFE) via electropolymerization. We have characterized these polyvanillin-carbon nanotube (PVN-MWNT) composite films with techniques including scanning electron microscopy (SEM), infrared spectroscopy (IR) and voltammetry. These investigations showed that the films have a uniform porous nanostructure with a large surface area. This PVN-MWNT composite-modified CFE (PVN-MWNT/CFE) exhibited a sensitive response to the electrochemical oxidation of nitrite. Under optimal working conditions, the oxidation peak current of nitrite linearly increased with its concentration in the range of 0.2 {mu}M-3.1 mM, with the system exhibiting a lower detection limit of 50 nM (S/N = 3). We successfully applied the PVN-MWNT/CFE system to the determination of nitrite from lake water. The efficient recovery of nitrite indicated that this electrode was able to detect nitrite in real samples.

Full Text Available The purpose of this paper is to evaluate the electrochemical behaviour of corrosion inhibition of the copper-carbon steel galvanic couple (Cu-CS, exposed to reverse osmosis water (RO used for rinsing of heat exchangers for heavy duty machinery, during manufacture. Molybdate and nitrite salts were utilized to evaluate the inhibition behaviour under galvanic couple conditions. Cu-CS couple was used as working electrodes to measure open circuit potential (OCP, potentiodynamic polarization (PP, and electrochemical impedance spectroscopy (EIS. The surface conditions were characterized by scanning electron microscopy (SEM and electron dispersive X-ray spectroscopy (EDS. The most effective concentration ratio between molybdate and nitrite corrosion inhibitors was determined. The morphological study indicated molybdate deposition on the anodic sites of the galvanic couple. The design of molybdate-based corrosion inhibitor developed in the present work should be applied to control galvanic corrosion of the Cu-CS couple during cleaning in the manufacture of heat exchangers.

Full Text Available Nitrogen is a main contaminant of wastewater worldwide. Novel processes for nitrogen removal have been developed over the last several decades. One of these is the partial nitritation process. This process includes the oxidation of ammonium to nitrite without the generation of nitrate. The partial nitritation process has several advantages over traditional nitrification-denitrification processes for nitrogen removal from wastewaters. In addition, partial nitritation is required for anammox elimination of nitrogen from wastewater. Partial nitritation is affected by operational conditions and substances present in the influent, such as quinolone antibiotics. In this review, the impact that several operational conditions, such as temperature, pH, dissolved oxygen concentration, hydraulic retention time and solids retention time, have over the partial nitritation process is covered. The effect of quinolone antibiotics and other emerging contaminants are discussed. Finally, future perspectives for the partial nitritation process are commented upon.

The effect of the initial nitrite concentration on the growth of S. platensis was experimentally investigated using the medium added to the sodium nitrite of 0.0-1.6 kg{center_dot}m{sup -3}. When the initial nitrite concentration is smaller than 0.8 kg{center_dot}m{sup -3}, the growth rate is greater than that without nitrite. The specific growth rate had the maximum value and is approximately 1.3 times that of a standard medium when the range of the initial nitrite concentration is 0.3-0.4 kg{center_dot}m{sup -3}. When the initial nitrite concentration is greater than 0.8 kg{center_dot}m{sup -3}, the growth rate was smaller than that of a standard medium due to nitrite inhibition. (author)

On September 9,Liangshan Mining Company’s 100,000 tons/year cathode copper project kicked off.It is another key project of the company following the successful launch of the 100,000 tons/year anode copper project.Based on ISA copper smelting technology of the largest open-cast copper mine in southwest China,

The present study focused on cupric sulphate and cupric nitrate uptake in Typha latifolia and the impact of these copper species on the plant's detoxification capacity. When the plants were exposed to 10, 50 and 100 μM cupric sulphate or cupric nitrate, copper accumulation in T. latifolia roots and shoots increased with rising concentration of the salts. Shoot to root ratios differed significantly depending on the form of copper supplementation, e.g. if it was added as cupric (II) sulphate or cupric (II) nitrate. After incubation with 100 μM of cupric sulphate, up to 450 mg Cu/kg fresh weight (FW) was accumulated, whereas the same concentration of cupric nitrate resulted in accumulation of 580 mg/kg FW. Furthermore, significant differences in the activity of some antioxidative enzymes in Typha roots compared to the shoots, which are essential in the plant's reaction to cope with metal stress, were observed. The activity of peroxidase (POX) in roots was increased at intermediate concentrations (10 and 50 μM) of CuSO4, whereas it was inhibited at the same Cu(NO3)2 concentrations. Ascorbate peroxidase (APOX) and dehydroascorbate reductase (DHAR) increased their enzyme activity intensely, which may be an indication for copper toxicity in T. latifolia plants. Besides, fluorodifen conjugation by glutathione S-transferases (GSTs) was increased up to sixfold, especially in roots.

.... Nitrate is a natural compound found in fruits and vegetables and when secreted in saliva, is reduced to nitrite through bacterial respiration and subsequently reduced to nitric oxide in acidic condition...

and phenanthrene, a polycyclic aromatic hydrocarbon. Analyses of different biomarkers in the treated fish indicated significant increase in the metabolism of phenanthrene as a result of exposure to nitrite. For example, the activity of the biotransformation enzyme...

Full Text Available The last decades have witnessed a steady increase of the social and political awareness for the need of monitoring and controlling environmental and industrial processes. In the case of nitrite ion, due to its potential toxicity for human health, the European Union has recently implemented a number of rules to restrict its level in drinking waters and food products. Although several analytical protocols have been proposed for nitrite quantification, none of them enable a reliable and quick analysis of complex samples. An alternative approach relies on the construction of biosensing devices using stable enzymes, with both high activity and specificity for nitrite. In this paper we review the current state-of-the-art in the field of electrochemical and optical biosensors using nitrite reducing enzymes as biorecognition elements and discuss the opportunities and challenges in this emerging market.

Nitrite-oxidizing bacteria (NOB) catalyze the second step of nitrification, a major process of the biogeochemical nitrogen cycle, but the recognized diversity of this guild is surprisingly low and only two bacterial phyla contain known NOB...

Nitrospira are a diverse group of nitrite-oxidizing bacteria and among the environmentally most widespread nitrifiers. However, they remain scarcely studied and mostly uncultured. Based on genomic and experimental data from Nitrospira moscoviensis representing the ubiquitous Nitrospira lineage II...

Nitric oxide (NO) has numerous important functions in the kidney, and long-term blockage of nitric oxide synthases in rats by L-NAME results in severe hypertension and progressive kidney damage. On the other hand, NO production seems to be low in patients with chronic kidney disease (CKD), and NO deficiency may play a role in CKD progression. In this review, we summarized the mechanisms of amelioration of renal injury induced by L-NAME treated rats by treatment of nitrite. First, we demonstrate whether orally-administrated nitrite-derived NO can shift to the circulation. When 3mg/kg body weight Na(15)NO(2) was orally administered to rats, an apparent EPR signal derived from Hb(15)NO (A(z)=23.4 gauss) appeared in the blood, indicating that orally ingested nitrite can be a source of NO in vivo. Next, in order to clarify the capacity of nitrite to prevent renal disease, we administered low-dose nitrite (LDN: 0.1mg of sodium nitrite in 1L of drinking water), medium-dose nitrite (MDN: 1mg sodium nitrite/L, which corresponds to the amount of nitrite ingested by vegetarians), or high-dose nitrite (HDN: 10mg sodium nitrite/L) to rats simultaneously with L-NAME (1 g l-NAME/L) for 8 weeks, then examined the blood NO level as a hemoglobin-NO adduct (iron-nitrosyl-hemoglobin) using electron paramagnetic resonance spectroscopy, urinary protein excretion, and renal histological changes at the end of the experiment. It was found that oral administration of MDN and HDN but not LDN increased the blood iron-nitrosyl-hemoglobin concentration to the normal level, ameliorated the L-NAME-induced proteinuria, and reduced renal histological damage. The findings demonstrate that chronic administration of a mid-level dietary dose of nitrite restores the circulating iron-nitrosyl-hemoglobin levels reduced by L-NAME and that maintenance of the circulating iron-nitrosyl-hemoglobin level in a controlled range protects against L-NAME-induced renal injury. Taking these findings together, we

In biological air filters ammonia is removed due to the action of Ammonia Oxidizing Bacteria (AOB) resulting in nitrite accumulation exceeding 100 mM. Among filters treating exhaust air from pig facilities successful establishment of Nitrite Oxidizing Bacteria (NOB) sometimes occurs, resulting...... in accumulation of nitrate rather than nitrite and a significant decline in pH. As a consequence, ammonia is removed more efficiently, but heterotrophic oxidation of odorous compounds might be inhibited. To identify the controlling mechanisms of nitrite oxidation, full-scale biological air filters were...... investigated applying a broad in situ approach. Bacterial distribution and chemical gradients were examined at both a macro scale from filter inlet towards outlet and on a micro scale within the intact biofilm, applying Fluorescence in situ Hybridization (FISH), electrochemical microsensors, and chemical...

Harvesting biohydrogen from inhibiting wastewaters is of practical interest since the toxicity of compounds in a wastewater stream commonly prevents the bioenergy content being recovered. The isolated Clostridium sp. R1 is utilized to degrade cellobiose in sulfide or nitrite-containing medium for biohydrogen production. The strain can effectively degrade cellobiose free of severe inhibitory effects at up to 200 mgl(-1) sulfide or to 5 mgl(-1) nitrite, yielding hydrogen at >2.0 mol H2 mol(-1) cellobiose. Principal metabolites of cellobiose fermentation are acetate and butyrate, with the concentration of the former increases with increasing sulfide and nitrite concentrations. The isolated strain can yield hydrogen from cellobiose in sulfide-laden wastewaters. However, the present of nitrite significantly limit the efficiency of the biohydrogen harvesting process.

The transition metal copper is an essential cofactor for many redox-active enzymes, but excessive copper can generate toxic reactive oxygen species. Copper homeostasis is maintained by highly conserved proteins, to balance copper uptake, distribution and export on the systemic and cellular level. Th

Copper tubes having diameters between about 100 and about 200 nm have been fabricated by electrodeposition of copper into the pores of alumina nanopore membranes. Copper nanotubes are under consideration as alternatives to copper nanorods and nanowires for applications involving thermal and/or electrical contacts, wherein the greater specific areas of nanotubes could afford lower effective thermal and/or electrical resistivities. Heretofore, copper nanorods and nanowires have been fabricated by a combination of electrodeposition and a conventional expensive lithographic process. The present electrodeposition-based process for fabricating copper nanotubes costs less and enables production of copper nanotubes at greater rate.

As nitrogen discharge limits are becoming more stringent, short-cut nitrogen systems and tertiary nitrogen polishing steps are gaining popularity. For partial nitritation or nitritation-denitritation systems, anaerobic ammonia oxidation (anammox) polishing may be feasible to remove residual ammonia and nitrite from the effluent. Nitrogen polishing of mainstream nitritation-denitritation system effluent via anammox was studied at 25°C in a fully anoxic moving bed bioreactor (MBBR) (V = 0.45 m(3) ) over 385 days. Unlike other anammox based processes, a very fast startup of anammox MBBR was demonstrated, despite nitrite limited feeding conditions (influent nitrite = 0.7 ± 0.59 mgN/L, ammonia = 6.13 ± 2.86 mgN/L, nitrate = 3.41 ± 1.92 mgN/L). The nitrogen removal performance was very stable within a wide range of nitrogen inputs. Anammox bacteria (AMX) activity up to 1 gN/m(2) /d was observed which is comparable to other biofilm-based systems. It is generally believed that nitrate production limits nitrogen removal through AMX metabolism. However, in this study, anammox MBBR demonstrated ammonia, nitrite, and nitrate removal at limited chemical oxygen demand (COD) availability. AMX and heterotrophs contributed to 0.68 ± 0.17 and 0.32 ± 0.17 of TIN removal, respectively. It was speculated that nitrogen removal might be aided by denitratation which could be due to heterotrophs or the recently discovered ability for AMX to use short-chain fatty acids to reduce nitrate to nitrite. This study demonstrates the feasibility of anammox nitrogen polishing in an MBBR is possible for nitritation-denitration systems.

Responses to glyceryl trinitrate/nitroglycerin (GTN), S-nitrosoglutathione (GSNO), and sodium nitrite were compared in the intact chest rat. The iv injections of GTN, sodium nitrite, and GSNO produced dose-dependent decreases in pulmonary and systemic arterial pressures. In as much as cardiac output was not reduced, the decreases in pulmonary and systemic arterial pressures indicate that GTN, sodium nitrite, and GSNO have significant vasodilator activity in the pulmonary and systemic vascular beds in the rat. Responses to GTN were attenuated by cyanamide, but not allopurinol, whereas responses to nitrite formed by the metabolism of GTN were attenuated by allopurinol and cyanamide. The results with allopurinol and cyanamide suggest that only mitochondrial aldehyde dehydrogenase is involved in the bioactivation of GTN, sodium nitrite, and GSNO, whereas both pathways are involved in the bioactivation of nitrite anion in the intact rat. The comparison of vasodilator activity indicates that GSNO and GTN are more than 1000-fold more potent than sodium nitrite in decreasing pulmonary and systemic arterial pressures in the rat. Following administration of 1H-[1,2,4]-oxadizaolo[4,3-]quinoxaline-1-one (ODQ), responses to GTN were significantly attenuated, indicating that responses are mediated by the activation of soluble guanylyl cyclase. These data suggest that the reduction of nitrite to nitric oxide formed from the metabolism of GTN, cannot account for the vasodilator activity of GTN in the intact rat and that another mechanism; perhaps the formation of an S-NO, may mediate the vasodilator response to GTN in this species.

Eosinophil peroxidase is a haem enzyme of eosinophils that is implicated in oxidative tissue injury in asthma. It uses hydrogen peroxide to oxidize thiocyanate and bromide to their respective hypohalous acids. Nitrite is also a substrate for eosinophil peroxidase. We have investigated the mechanisms by which the enzyme oxidizes nitrite. Nitrite was very effective at inhibiting hypothiocyanous acid ('cyanosulphenic acid') and hypobromous acid production. Spectral studies showed that nitrite reduced the enzyme to its compound II form, which is a redox intermediate containing Fe(IV) in the haem active site. Compound II does not oxidize thiocyanate or bromide. These results demonstrate that nitrite is readily oxidized by compound I, which contains Fe(V) at the active site. However, it reacts more slowly with compound II. The observed rate constant for reduction of compound II by nitrite was determined to be 5.6x10(3) M(-1) x s(-1). Eosinophils were at least 4-fold more effective at promoting nitration of a heptapeptide than neutrophils. This result is explained by our finding that nitrite reacts 10-fold faster with compound II of eosinophil peroxidase than with the analogous redox intermediate of myeloperoxidase. Nitration by eosinophils was increased 3-fold by superoxide dismutase, which indicates that superoxide interferes with nitration. We propose that at sites of eosinophilic inflammation, low concentrations of nitrite will retard oxidant production by eosinophil peroxidase, whereas at higher concentrations nitrogen dioxide will be a major oxidant formed by these cells. The efficiency of protein nitration will be decreased by the diffusion-controlled reaction of superoxide with nitrogen dioxide.

Nitrification, the sequential aerobic oxidation of ammonia via nitrite to nitrate, is a key process of the biogeochemical nitrogen cycle and catalyzed by two aerobic microbial guilds (nitrifiers): ammonia oxidizers and nitrite-oxidizing bacteria (NOB). NOB are generally considered as metabolically restricted and dependent on ammonia oxidizers. Here, we report that, surprisingly, key NOB of many ecosystems (Nitrospira) convert urea, an important ammonia source in nature, to ammonia and CO2. Th...

Nitrite is an important reservoir of nitric oxide activity in the plasma and cells. Using a biomimetic model, we demonstrate the conversion of zinc-bound nitrite in the tris(pyrazolyl)borate complex (iPr2)TpZn(NO2) to the corresponding S-nitrosothiol RSNO and zinc thiolate (iPr2)TpZn-SR via reaction with thiols H-SR. Decomposition of the S-nitrosothiol formed releases nitric oxide gas.

Cyclic polarization scans were performed using A-537 carbon steel in simulated washed precipitate solutions of various nitrite and nitrate concentrations. The results of this study indicate that nitrate is an aggressive anion in washed precipitate. Furthermore, a quantitative linear log-log relationship between the minimum effective nitrite concentration and the nitrate concentration was established for washed precipitate with other ions at their average compositions.

A critical review is presented of the possibility of stress corrosion cracking (SCC) of copper canisters in a deep geological repository in the Fennoscandian Shield. Each of the four main mechanisms proposed for the SCC of pure copper are reviewed and the required conditions for cracking compared with the expected environmental and mechanical loading conditions within the repository. Other possible mechanisms are also considered, as are recent studies specifically directed towards the SCC of copper canisters. The aim of the review is to determine if and when during the evolution of the repository environment copper canisters might be susceptible to SCC. Mechanisms that require a degree of oxidation or dissolution are only possible whilst oxidant is present in the repository and then only if other environmental and mechanical loading conditions are satisfied. These constraints are found to limit the period during which the canisters could be susceptible to cracking via film rupture (slip dissolution) or tarnish rupture mechanisms to the first few years after deposition of the canisters, at which time there will be insufficient SCC agent (ammonia, acetate, or nitrite) to support cracking. During the anaerobic phase, the supply of sulphide ions to the free surface will be transport limited by diffusion through the highly compacted bentonite. Therefore, no HS. will enter the crack and cracking by either of these mechanisms during the long term anaerobic phase is not feasible. Cracking via the film-induced cleavage mechanism requires a surface film of specific properties, most often associated with a nano porous structure. Slow rates of dissolution characteristic of processes in the repository will tend to coarsen any nano porous layer. Under some circumstances, a cuprous oxide film could support film-induced cleavage, but there is no evidence that this mechanism would operate in the presence of sulphide during the long-term anaerobic period because copper sulphide

Full Text Available Aspects concerning nitrate and nitrite pollution of groundwaters. Water is a basic natural resource for the good functioning of all thebiological processes in nature. It is very important for life and for the developmentof human activities. The quality of the ground water has begun to degrade moreand more, as a result of the physical, chemical and bacteriological changes.Nitrogen compounds pollution of the underground has increased lately. This hasbeen caused by the excessive and irrational use of nitrogen derived fertilizers, bythe wrong storage of the dejections resulted from zootechnical processes and byother chemical substances discharged into water. Samples were collected fromdifferent wells in order to check whether the well water was drinkable. The resultof the test revealed the existence of high concentrations of nitrates as well asvalues exceeding normal microbiological parameters. The value recorded in thetown of Segarcea, the county of Dolj, showed extremely high concentrations ofnitrates of the drinking water in the wells. Thus, Segarcea is the town with thegreatest number of contaminated wells in the country.

Oxidative stress is usually considered as an important factor to the pathogenesis of various diseases. Peroxynitrite (ONOO(-)) and hypochlorite (OCl(-)) are formed in immune cells as a part of the innate host defense system, but excessive reactive oxygen species generation can cause progressive inflammation and tissue damage. It has been proven that through mediating nitric oxide (NO) homeostasis, inorganic nitrite (NO2(-)) shows organ-protective effects on oxidative stress and inflammation. However, the effects of NO2(-) on the function of immune cells were still not clear. The potential role of NO2(-) in modulating ONOO(-) and OCl(-) generation in neutrophil cells was investigated in this study. As an immune cell activator, lipopolysaccharide (LPS) increased both ONOO(-) and OCl(-) production in neutrophils, which was significantly attenuated by NO2(-). NO2(-) reduced superoxide (O2(·-)) generation via a NO-dependent mechanism and increased NO formation in activated neutrophils, suggesting a crucial role of O2(·-) in NO2(-)-mediated reduction of ONOO(-). Moreover, the reduced effect of NO2(-) on OCl(-) production was attributed to that NO2(-) reduced H2O2 production in activated neutrophils without influencing the release of myeloperoxidase (MPO), thus limiting OCl(-) production by MPO/H2O2 system. Therefore, NO2(-) attenuates ONOO(-) and OCl(-) formation in activated neutrophils, opening a new direction to modulate the inflammatory response.

Copper is one of the essential trace elements. It is part of a number of enzymes. Deficiency of the element is manifested by impaired haematopoesis, bone metabolism, disorders of the digestive, cardiovascular and nervous system. Deficiency occurs in particular in patients suffering from malnutrition, malabsorption, great copper losses during administration of penicillamine. Sporadically copper intoxications are described (suicidal intentions or accidental ingestion of beverages with a high copper content). Acute exposure to copper containing dust is manifested by metal fume fever. Copper salts can produce local inflammations. Wilson's disease is associated with inborn impaired copper metabolism. In dialyzed patients possible contaminations of the dialyzate with copper must be foreseen as well as the possible release of copper from some dialyzation membranes. With the increasing amount of copper in the environment it is essential to monitor the contamination of the environment.

The correct levels of deoxyribonucleotide triphosphates and their relative abundance are important to maintain genomic integrity. Ribonucleotide reductase (RNR) regulation is complex and multifaceted. RNR is regulated allosterically by two nucleotide-binding sites, by transcriptional control, and...

To help learn how phytopathogens feed from their hosts, genes for nutrient transporters from the hemibiotrophic potato and tomato pest Phytophthora infestans were annotated. This identified 453 genes from 19 families. Comparisons with a necrotrophic oomycete, Pythium ultimum var. ultimum, and a hemibiotrophic fungus, Magnaporthe oryzae, revealed diversity in the size of some families although a similar fraction of genes encoded transporters. RNA-seq of infected potato tubers, tomato leaves, and several artificial media revealed that 56 and 207 transporters from P. infestans were significantly up- or down-regulated, respectively, during early infection timepoints of leaves or tubers versus media. About 17 were up-regulated >4-fold in both leaves and tubers compared to media and expressed primarily in the biotrophic stage. The transcription pattern of many genes was host-organ specific. For example, the mRNA level of a nitrate transporter (NRT) was about 100-fold higher during mid-infection in leaves, which are nitrate-rich, than in tubers and three types of artificial media, which are nitrate-poor. The NRT gene is physically linked with genes encoding nitrate reductase (NR) and nitritereductase (NiR), which mobilize nitrate into ammonium and amino acids. All three genes were coregulated. For example, the three genes were expressed primarily at mid-stage infection timepoints in both potato and tomato leaves, but showed little expression in potato tubers. Transformants down-regulated for all three genes were generated by DNA-directed RNAi, with silencing spreading from the NR target to the flanking NRT and NiR genes. The silenced strains were nonpathogenic on leaves but colonized tubers. We propose that the nitrate assimilation genes play roles both in obtaining nitrogen for amino acid biosynthesis and protecting P. infestans from natural or fertilization-induced nitrate and nitrite toxicity. PMID:27936244

This study investigated the effects of applying different doses of gamma radiation (0, 10 and 20 kGy) on Clostridium botulinum spores (107 spores/g) inoculated into mortadellas with different nitrite contents (0, 150 and 300 ppm). We also evaluated the order of application of heat (cooking) and irradiation processing. The products were evaluated for survival of C. botulinum, pH, water activity (Aw), redox potential (Eh) and residual nitrite content. In the non-irradiated raw batters, almost all spores could be recovered when no nitrite was added and only half was recovered with the addition of 150 ppm of nitrite. The use of 150 ppm of nitrite was able to inhibit the germination or growth of C. botulinum in non-irradiated cooked mortadellas after 48 h of processing. However, after 30 days of chilling storage (4 °C), it was possible to recover 105 UFC/g of this microorganism. The gamma irradiation (>10 kGy) had a positive effect on the inactivation of C. botulinum in mortadellas, independent of the sodium nitrite level used and the cooking/irradiation processing order.

A mathematical model based on Activated Sludge Model No. 3 (International Water Association, London) and laboratory-scale experiments were used to investigate ammonia conversion by nitrification in a sequencing batch reactor (SBR). The purpose of the study was to assess the effect of dissolved oxygen concentration on nitrite accumulation in the SBR. As the dissolved oxygen concentration in the SBR depends on the balance between oxygen consumption and oxygen transfer rates, ammonium conversion was measured for different air flowrate values to obtain different dissolved oxygen concentration profiles during the cycle. The ammonia concentration in the feeding medium was 500 mg ammonium as nitrogen (N-NH4(+))/L, and the maximum nitrite concentration achieved during a cycle was approximately 50 mg nitrite as nitrogen (N-NO2)/L. The air flow supplied to the reactor was identified as a suitable parameter to control nitrite accumulation in the SBR. This identification was carried out based on experimental results and simulation with a calibrated model. At a low value of the volumetric mass-transfer coefficient (kLa), the maximum nitrite concentration achieved during a cycle depends strongly on k(L)a, whereas, at a high value of k(L)a, the maximum nitrite concentration was practically independent of kL(a).

The sequential statistical experimental design (Plackett-Burman, factorial, response surface and steepest ascent experiment) was applied to optimize the culture medium of nitrite oxidizing bacteria for improving the nitrite oxidizing rate. Estimated optimum medium composition of the nitrite oxidizing rate was as follows: NaHCO3, 1.86gl(-1); NaNO2, 2.04gl(-1); Na2CO3, 0.2gl(-1); NaCl, 0.2gl(-1); KH2PO4, 0.1gl(-1); MgSO4 x7H2O, 0.1gl(-1); and FeSO4 x 7H2O, 0.01gl(-1). The nitrite oxidizing rate was increased by 48.0% and reached a maximum at 859.5+/-8.4mgNO2-N/gMLSS.d as compared to 580.7+/-25.8mgNO2-N/gMLSS x d. In the field trial, 50L of nitrite oxidizing bacteria concentrate (1.99gVSS/L) with 850mgNO2-N/gMLSS x d were added to 0.6ha of the aquaculture water. Nitrite level in all treated ponds remained very low compared to the steady increase observed in all of the control ponds during 7 days.

Full Text Available In April, 2013, a Suzhou Hospital reported a nitrite intoxication patient in coma as well as 2 family members with the similar symptom 5 days ahead. We investigated the event to identify the cause, source and possible pollution ways of the contamination.We defined case as any person living in YSHY community who has cyanosis and with at least one of the following symptoms: dizziness, headache, fatigue, tachycardia, drowsiness, nausea, vomiting, abdominal pain or diar-rhea during April 15 to April 25, 2013. We searched for case by interviewing community residents and reviewing clinics' medical records; information was then retrospectively collected on the patient's food history, cooking procedures and food sources.We identified 3 nitrite intoxication cases, 1 male and 2 female from a family. The interval time between dinner and onset was < 1 hour. Retrospective survey showed 'sugar stir and mix asparagus' on April 17 and 'scrambled asparagus' on April 21 were suspected foods. Both suspected dishes had 'sugar' added, sourced from a clean-up of a neighboring rental house. Nitrite was detected in a vomitus sample, the 'sugar' and two leftover food samples.This family clustered nitrite intoxication was induced by using unidentified nitrite as sugar to cook dishes. We recommend sodium nitrite should be dyed with bright colors to avoid mistaking it for plain salt or sugar, health departments strengthen food hygiene propaganda to improve people's recognition of food safety, and to alert them the dangerous of eating unidentified or unknown source food.

Full Text Available The effect of lycopene on serum nitrate-nitrite levels was investigated in diabetic rats. In this investigation, 28 Wistar rats were divided into 4 groups, each of seven rats. These groups were control group, diabetes group, diabetes-lycopene group and lycopene group. The concentration of nitrite and nitrate was detected at high levels in diabetes group, diabetes-lycopene group and lycopene group as compared with the control group (P<0.05. Especially, the increase in the levels of nitrate in diabetes group and lycopene group was statistically significant when compared with diabetes-lycopene group and control group (P<0.05. In addition, we also determined the proportion of nitrite/nitrate for nitric oxide radical formation. Therefore, it is important to investigate the recovery and stability of nitrite and nitrate in samples. As a result of this study, it was observed that the amounts of nitrate and nitrite increased due to oxidative stress in diabetes and also application of antioxidant lycopene caused an increase in the amounts of nitrate and nitrite levels.

Pd-based catalyst treatment represents an emerging technology that shows promise to remove nitrate and nitrite from drinking water. In this work we use vapor-grown carbon nanofiber (CNF) supports in order to explore the effects of Pd nanoparticle size and interior versus exterior loading on nitrite reduction activity and selectivity (i.e., dinitrogen over ammonia production). Results show that nitrite reduction activity increases by 3.1-fold and selectivity decreases by 8.0-fold, with decreasing Pd nanoparticle size from 1.4 to 9.6 nm. Both activity and selectivity are not significantly influenced by Pd interior versus exterior CNF loading. Consequently, turnover frequencies (TOFs) among all CNF catalysts are similar, suggesting nitrite reduction is not sensitive to Pd location on CNFs nor Pd structure. CNF-based catalysts compare favorably to conventional Pd catalysts (i.e., Pd on activated carbon or alumina) with respect to nitrite reduction activity and selectivity, and they maintain activity over multiple reduction cycles. Hence, our results suggest new insights that an optimum Pd nanoparticle size on CNFs balances faster kinetics with lower ammonia production, that catalysts can be tailored at the nanoscale to improve catalytic performance for nitrite, and that CNFs hold promise as highly effective catalyst supports in drinking water treatment.

Full Text Available Hypertension is a common disease that includes oxidative stress as a major feature, and oxidative stress impairs physiological nitric oxide (NO activity promoting cardiovascular pathophysiological mechanisms. While inorganic nitrite and nitrate are now recognized as relevant sources of NO after their bioactivation by enzymatic and non-enzymatic pathways, thus lowering blood pressure, mounting evidence suggests that sodium nitrite also exerts antioxidant effects. Here we show for the first time that sodium nitrite exerts consistent systemic and vascular antioxidant and antihypertensive effects in the deoxycorticosterone-salt (DOCA-salt hypertension model. This is particularly important because increased oxidative stress plays a major role in the DOCA-salt hypertension model, which is less dependent on activation of the renin-angiotensin system than other hypertension models. Indeed, antihypertensive effects of oral nitrite were associated with increased plasma nitrite and nitrate concentrations, and completely blunted hypertension-induced increases in plasma 8-isoprostane and lipid peroxide levels, in vascular reactive oxygen species, in vascular NADPH oxidase activity, and in vascular xanthine oxidoreductase activity. Together, these findings provide evidence that the oral administration of sodium nitrite consistently decreases the blood pressure in association with major antioxidant effects in experimental hypertension.

This study developed a simple numerical model for nitrogen removal in biofilters, which was designed to enhance simultaneous nitritation and anaerobic ammonium oxidation (anammox). It is the first attempt to simulate anammox together with two-step nitrification in natural treatment systems, which may have different kinetic parameters and temperature effects from conventional bioreactors. Prediction accuracy was improved by adjusting kinetic coefficients over the startup period of the biofilters. The maximum rates of nitritation and nitrite oxidation increased linearly over time during the startup period. Simulations confirmed successful enhancement of simultaneous nitritation and anammox (SNA) in the biofilters, with anammox contributing 35% of ammonium removal. Effluent ammonium concentration was affected by influent ammonium concentration and the maximum nitritation rate, and was insensitive to the maximum nitrite oxidation rate and anammox substrate factor. Ammonium removal via SNA was likely limited by biomass of aerobic ammonia oxidizing bacteria in the biofilters. The developed model is a promising tool for studying the dynamics of nitrogen removal processes including SNA in natural treatment systems.

Full Text Available Background: In April 2013, a hospital in Suzhou City notified authorities of a patient with nitrite poisoning with two other family members who had similar toxic symptoms five days prior. We investigated the event to identify the cause, source and possible route of contamination. Methods: A case was defined as any person living in the Yang Shan Hua Yuan community who had been diagnosed with cyanoderma and food poisoning symptoms from 15 to 25 April 2013. Active case finding was conducted by interviewing community residents and reviewing medical records from local clinics; information was then retrospectively collected on the patient’s food history, cooking procedures and food sources. Results: We identified three nitrite poisoning cases, one male and two females, from the same family. The time between dinner and onset of illness was less than an a hour. A retrospective survey showed that a substance presumed to be sugar mixed with asparagus on 17 April and with stir-fried asparagus on 21 April wasthe suspected contaminant. The presumed sugar came from a clean-up of a neighbouring rental house. Nitrite was detected in a vomitus sample, the sugar substance and two leftover food samples. Conclusion: This family cluster of nitrite poisoning resulted from the mistaken use of nitrite as sugar to cook dishes. We recommend that sodium nitrite be dyed a bright colour to prevent such a mistake and that health departments strengthen food hygiene education to alert people about the danger of eating unidentified food from an unknown source.

Lactobacillus fermentum was substituted for nitrite to produce cured pink color in a Chinese-style sausage. Treatments included inoculations (10(4), 10(6), and 10(8)CFU/g meat) followed by fermentation at 30°C for 8h and then at 4°C for 16h. Control sausage (with sodium nitrite, 60mg/kg meat) was cured at 4°C for 24h without L. fermentum. The UV-Vis spectra of pigment extract from L. fermentum-treated sausage were identical to that of nitrosylmyoglobin (NO-Mb) formed in nitrite-treated control. The NO-Mb concentration and the colorimetric a(∗) value of sausage treated with 10(8)CFU/g meat of L. fermentum essentially replicated those in nitrite-cured meat. Free amino acid content in sausage treated with L. fermentum was greater and the pH slightly lower compared with the nitrite-cured control sample. This study showed that L. fermentum has the potential to substitute for nitrite in the sausage production.

The present paper seeks to develop a simple method for the spectrophotometric determination of nitrite in soil and water samples and also measure optimum reaction conditions along with other analytical parameters. The method is based on the diazotization-coupling reaction of nitrite with cefixime and 1-naphthylamine in an acidic solution (Griess reaction). The final product that is an azo dye has an orange color with maximum absorption at 360 nm which Beer's Law is obeyed over the concentration range 0.02-15.00 mg L-1 of nitrite. Optimal conditions of the variables affecting the reaction were obtained by central composite design (CCD). A detection limit of 4.3 × 10-3 mg L-1 was obtained for determination of nitrite by the proposed method. The proposed method was successfully applied to determine nitrite in soil and water samples. The molar absorptivity of the product of the reaction and RSD in determination of nitrite in real samples are 4.1 × 103 (L mol-1 cm-1) and lower than 10%, respectively.

Nitric oxide (NO) is generated endogenously by NO synthases to regulate a number of physiological processes including cardiovascular and metabolic functions. A decrease in the production and bioavailability of NO is a hallmark of many major chronic diseases including hypertension, ischaemia-reperfusion injury, atherosclerosis and diabetes. This NO deficiency is mainly caused by dysfunctional NO synthases and increased scavenging of NO by the formation of reactive oxygen species. Inorganic nitrate and nitrite are emerging as substrates for in vivo NO synthase-independent formation of NO bioactivity. These anions are oxidation products of endogenous NO generation and are also present in the diet, with green leafy vegetables having a high nitrate content. The effects of nitrate and nitrite are diverse and include vasodilatation, improved endothelial function, enhanced mitochondrial efficiency and reduced generation of reactive oxygen species. Administration of nitrate or nitrite in animal models of cardiovascular disease shows promising results, and clinical trials are currently ongoing to investigate the therapeutic potential of nitrate and nitrite in hypertension, pulmonary hypertension, peripheral artery disease and myocardial infarction. In addition, the nutritional aspects of the nitrate-nitrite-NO pathway are interesting as diets suggested to protect against cardiovascular disease, such as the Mediterranean diet, are especially high in nitrate. Here, we discuss the potential therapeutic opportunities for nitrate and nitrite in prevention and treatment of cardiovascular and metabolic diseases.

Recent evidence suggests that the reaction of nitrite with deoxygenated hemoglobin and myoglobin contributes to the generation of nitric oxide and S-nitrosothiols in vivo under conditions of low oxygen availability. We have investigated whether ferrous neuroglobin and cytoglobin, the two hexacoor......Recent evidence suggests that the reaction of nitrite with deoxygenated hemoglobin and myoglobin contributes to the generation of nitric oxide and S-nitrosothiols in vivo under conditions of low oxygen availability. We have investigated whether ferrous neuroglobin and cytoglobin, the two...... hexacoordinate globins from vertebrates expressed in brain and in a variety of tissues, respectively, also react with nitrite under anaerobic conditions. Using absorption spectroscopy, we find that ferrous neuroglobin and nitrite react with a second-order rate constant similar to that of myoglobin, whereas...... the ferrous heme of cytoglobin does not react with nitrite. Deconvolution of absorbance spectra shows that, in the course of the reaction of neuroglobin with nitrite, ferric Fe(III) heme is generated in excess of nitrosyl Fe(II)-NO heme as due to the low affinity of ferrous neuroglobin for nitric oxide...

Many of the proteins that are candidates for bioenergetic pathways involved with sulfate respiration in Desulfovibrio spp. have been studied, but complete pathways and overall cell physiology remain to be resolved for many environmentally relevant conditions. In order to understand the metabolism of these microorganisms under adverse environmental conditions for improved bioremediation efforts, Desulfovibrio vulgaris Hildenborough was used as a model organism to study stress response to nitrite, an important intermediate in the nitrogen cycle. Previous physiological studies demonstrated that growth was inhibited by nitrite and that nitrite reduction was observed to be the primary mechanism of detoxification. Global transcriptional profiling with whole-genome microarrays revealed coordinated cascades of responses to nitrite in pathways of energy metabolism, nitrogen metabolism, oxidative stress response, and iron homeostasis. In agreement with previous observations, nitrite-stressed cells showed a decrease in the expression of genes encoding sulfate reduction functions in addition to respiratory oxidative phosphorylation and ATP synthase activity. Consequently, the stressed cells had decreased expression of the genes encoding ATP-dependent amino acid transporters and proteins involved in translation. Other genes up-regulated in response to nitrite include the genes in the Fur regulon, which is suggested to be involved in iron homeostasis, and genes in the Per regulon, which is predicted to be responsible for oxidative stress response.

The partial nitrification (PN) performance and the microbial community variations were evaluated in a sequencing batch reactor (SBR) for 172 days, with the stepwise elevation of ammonium concentration. Free ammonia (FA) and low dissolved oxygen inhibition of nitrite-oxidized bacteria (NOB) were used to achieve nitritation in the SBR. During the 172 days operation, the nitrogen loading rate of the SBR was finally raised to 3.6 kg N/m3/d corresponding the influent ammonium of 1500 mg/L, with the ammonium removal efficiency and nitrite accumulation rate were 94.12% and 83.54%, respectively, indicating that the syntrophic inhibition of FA and low dissolved oxygen contributed substantially to the stable nitrite accumulation. The results of the 16S rRNA high-throughput sequencing revealed that Nitrospira, the only nitrite-oxidizing bacteria in the system, were successively inhibited and eliminated, and the SBR reactor was dominated finally by Nitrosomonas, the ammonium-oxidizing bacteria, which had a relative abundance of 83%, indicating that the Nitrosomonas played the primary roles on the establishment and maintaining of nitritation. Followed by Nitrosomonas, Anaerolineae (7.02%) and Saprospira (1.86%) were the other mainly genera in the biomass. PMID:27762325

Full Text Available Iron and copper are essential for plants and are important for the function of a number of protein complexes involved in photosynthesis and respiration. As the molecular mechanisms that control uptake, trafficking and storage of these nutrients emerge, the importance of metalloreductase-catalyzed reactions in iron and copper metabolism has become clear. This review focuses on the FRO family of metalloreductases in plants and highlights new insights into the roles of FRO family members in metal homeostasis. Arabidopsis FRO2 was first identified as the ferric chelate reductase that reduces ferric iron-chelates at the root surface-rhizosphere interface. The resulting ferrous iron is subsequently transported across the plasma membrane of root epidermal cells by the ferrous iron transporter, IRT1. Recent work has shown that two other members of the FRO family (FRO4 and FRO5 function redundantly to reduce copper to facilitate its uptake from the soil. In addition, FROs appear to play important roles in subcellular compartmentalization of iron as FRO7 is known to contribute to delivery of iron to chloroplasts while mitochondrial family members FRO3 and FRO8 are hypothesized to influence mitochondrial metal ion homeostasis. Finally, recent studies have underscored the importance of plasma membrane-localized ferric reductase activity in leaves for photosynthetic efficiency. Taken together, these studies highlight a number of diverse roles for FROs in both iron and copper metabolism in plants.

Copper is an essential micronutrient involved in fundamental life processes that are conserved throughout all forms of life. The ability of copper to catalyze oxidation-reduction (redox) reactions, which can inadvertently lead to the production of reactive oxygen species (ROS), necessitates the tight homeostatic regulation of copper within the body. Many cancer types exhibit increased intratumoral copper and/or altered systemic copper distribution. The realization that copper serves as a limiting factor for multiple aspects of tumor progression, including growth, angiogenesis and metastasis, has prompted the development of copper-specific chelators as therapies to inhibit these processes. Another therapeutic approach utilizes specific ionophores that deliver copper to cells to increase intracellular copper levels. The therapeutic window between normal and cancerous cells when intracellular copper is forcibly increased, is the premise for the development of copper-ionophores endowed with anticancer properties. Also under investigation is the use of copper to replace platinum in coordination complexes currently used as mainstream chemotherapies. In comparison to platinum-based drugs, these promising copper coordination complexes may be more potent anticancer agents, with reduced toxicity toward normal cells and they may potentially circumvent the chemoresistance associated with recurrent platinum treatment. In addition, cancerous cells can adapt their copper homeostatic mechanisms to acquire resistance to conventional platinum-based drugs and certain copper coordination complexes can re-sensitize cancer cells to these drugs. This review will outline the biological importance of copper and copper homeostasis in mammalian cells, followed by a discussion of our current understanding of copper dysregulation in cancer, and the recent therapeutic advances using copper coordination complexes as anticancer agents.

Peroxiredoxin (Prx) constitutes a large family of enzymes found in microorganisms, animals, and plants, but the detection of the activities of Prx-linked hydroperoxide reductases (peroxiredoxin reductases) in cell extracts, and the purification based on peroxide reductase activity, have only been done in bacteria and Trypanosomatidae. A peroxiredoxin reductase (NADH oxidase) from a bacterium, Amphibacillus, displayed only poor activities in the presence of purified Prx from Saccharomyces or Synechocystis, while it is highly active in the presence of bacterial Prx. These results suggested that an enzyme system different from that in bacteria might exist for the reduction of Prx in yeast and cyanobacteria. Prx-linked hydroperoxide reductase activities were detected in cell extracts of Saccharomyces, Synechocystis, and Chlorella, and the enzyme activities of Saccharomyces and Chlorella were induced under vigorously aerated culture conditions and intensive light exposure conditions, respectively. Partial purification of Prx-linked peroxidase from the induced yeast cells indicated that the Prx-linked peroxidase system consists of two protein components, namely, thioredoxin and thioredoxin reductase. This finding is consistent with the previous report on its purification based on its protein protection activity against oxidation [Chae et al., J. Biol. Chem., 269, 27670-27678 (1994)]. In this study we have confirmed that Prx-linked peroxidase activity are widely distributed, not only in bacteria species and Trypanosomatidae, but also in yeast and photosynthetic microorganisms, and showed reconstitution of the activity from partially purified interspecies components.

To understand the effects of physicochemical factors on nitrite transformation by microalgae, a lipid-rich Chlorella with high nitrite tolerance was cultured with 8 mmol/l sodium nitrite as sole nitrogen source under different conditions. The results showed that nitrite transformation was mainly dependent on the metabolic activities of algal cells rather than oxidation of nitrite by dissolved oxygen. Light intensity, temperature, pH, NaHCO3 concentrations, and initial cell densities had significant effects on the rate of nitrite transformation. Single-factor experiments revealed that the optimum conditions for nitrite transformation were light intensity: 300 μmol/m(2); temperature: 30°C; pH: 7-8; NaHCO3 concentration: 2.0 g/l; and initial cell density: 0.15 g/l; and the highest nitrite transformation rate of 1.36 mmol/l/d was achieved. There was a positive correlation between nitrite transformation rate and the growth of Chlorella. The relationship between nitrite transformation rate (mg/l/d) and biomass productivity (g/l/d) could be described by the regression equation y = 61.3x (R(2) = 0.9665), meaning that 61.3 mg N element was assimilated by 1.0 g dry biomass on average, which indicated that the nitrite transformation is a process of consuming nitrite as nitrogen source by Chlorella. The results demonstrated that the Chlorella suspension was able to assimilate nitrite efficiently, which implied the feasibility of using flue gas for mass production of Chlorella without preliminary removal of NOX.

Full Text Available Diabetes mellitus (DM is a complex metabolic disorder arising from lack of insulin production or insulin resistance 1. DM is a leading cause of morbidity and mortality in the developed world, particularly from vascular complications such as atherothrombosis in the coronary vessels. Aldose reductase (AR [ALR2; EC 1.1.1.21], a key enzyme in the polyol pathway, catalyzes NADPH-dependent reduction of glucose to sorbitol, leading to excessive accumulation of intracellular reactive oxygen species (ROS in various tissues of DM including the heart, vasculature, neurons, eyes and kidneys. As an example, hyperglycemia through such polyol pathway induced oxidative stress, may have dual heart actions, on coronary blood vessel (atherothrombosis and myocardium (heart failure leading to severe morbidity and mortality (reviewed in 2. In cells cultured under high glucose conditions, many studies have demonstrated similar AR-dependent increases in ROS production, confirming AR as an important factor for the pathogenesis of many diabetic complications. Moreover, recent studies have shown that AR inhibitors may be able to prevent or delay the onset of cardiovascular complications such as ischemia/reperfusion injury, atherosclerosis and atherothrombosis. In this review, we will focus on describing pivotal roles of AR in the pathogenesis of cardiovascular diseases as well as other diabetic complications, and the potential use of AR inhibitors as an emerging therapeutic strategy in preventing DM complications.

Retinal microglia (RMG) are one of the major immune cells in charge of surveillance of inflammatory responses in the eye. In the absence of an inflammatory stimulus, RMG reside predominately in the ganglion layer and inner or outer plexiform layers. However, under stress RMG become activated and migrate into the inner nuclear layer (INL) or outer nuclear layer (ONL). Activated RMG in cell culture secrete pro-inflammatory cytokines in a manner sensitive to downregulation by aldose reductase inhibitors. In this study, we utilized CX3CR1(GFP) mice carrying AR mutant alleles to evaluate the role of AR on RMG activation and migration in vivo. When tested on an AR(WT) background, IP injection of LPS induced RMG activation and migration into the INL and ONL. However, this phenomenon was largely prevented by AR inhibitors or in AR null mice, or was exacerbated in transgenic mice that over-express AR. LPS-induced increases in ocular levels of TNF-α and CX3CL-1 in WT mice were substantially lower in AR null mice or were reduced by AR inhibitor treatment. These studies demonstrate that AR expression in RMG may contribute to the proinflammatory phenotypes common to various eye diseases such as uveitis and diabetic retinopathy.

Retinal microglia (RMG) are one of the major immune cells in charge of surveillance of inflammatory responses in the eye. In the absence of an inflammatory stimulus, RMG reside predominately in the ganglion layer and inner or outer plexiform layers. However, under stress RMG become activated and migrate into the inner nuclear layer (INL) or outer nuclear layer (ONL). Activated RMG in cell culture secrete pro-inflammatory cytokines in a manner sensitive to downregulation by aldose reductase inhibitors. In this study, we utilized CX3CR1{sup GFP} mice carrying AR mutant alleles to evaluate the role of AR on RMG activation and migration in vivo. When tested on an AR{sup WT} background, IP injection of LPS induced RMG activation and migration into the INL and ONL. However, this phenomenon was largely prevented by AR inhibitors or in AR null mice, or was exacerbated in transgenic mice that over-express AR. LPS-induced increases in ocular levels of TNF-α and CX3CL-1 in WT mice were substantially lower in AR null mice or were reduced by AR inhibitor treatment. These studies demonstrate that AR expression in RMG may contribute to the proinflammatory phenotypes common to various eye diseases such as uveitis and diabetic retinopathy. - Highlights: • AR inhibition prevents retinal microglial activation. • Endotoxin-induced ocular cytokine production is reduced in AR null mice. • Overexpression of AR spontaneously induces retinal microglial activation.

Full Text Available Folate is a vital B vitamin which is easily water-soluble. It is a natural source which is found in the herbal and animal foods. Folate has important duties in the human metabolism, one of them is the adjustment of the level of plasma homocysteine. Reduction in MTHFR (methylenetetrahydrofolate reductase,which is in charge of the metabolism of homocysteine activity affects the level of homocysteine. Therefore MTHFR is an important enzyme in folate metabolism. Some of the mutations occurring in the MTHFR gene is a risk factor for various diseases and may be caused the hyperhomocysteinemia or the homocystinuria, and they also may lead to metabolic problems. MTHFR is effective in the important pathways such as DNA synthesis, methylation reactions and synthesis of RNA. C677T and A1298C are the most commonly occurring polymorphisms in the gene of MTHFR. The frequency of these polymorphisms show differences in the populations. MTHFR, folate distribution, metabolism of homocysteine and S-adenosylmethionine, by the MTHFR methylation the genetic defects have the potential of affecting the risk of disease in the negative or positive way.

Full Text Available In the previous part, we presented the standard operating procedure (SOP of the microscopic observation drug susceptibility assay drug susceptibility testing (DST for Mycobacterium tuberculosis. The present SOP is devoted to another non-commercial culture and DST method known as nitrate reductase assay (NRA. As the name implies, the NRA detects the ability of M. tuberculosis to reduce nitrate to nitrite. In the assay, the presence of nitrite is detected by the addition of p-nitrobenzoate into the growth yield. The reaction is detected by the naked eye. The incorporation of drugs in the medium allows to use the test for DST, which can be interpreted with naked eyes. The identification and drug susceptibility results can be obtained in 2-3 weeks. This SOP document has been developed through the culture and DST subgroup of the STOP tuberculosis (TB Partnership New Diagnostic Working Group. It is intended for laboratories that would want to use or already using this rapid non-commercial method for culture identification and DST of M. tuberculosis, notably in resource-constraint settings in Asia and Africa.

These copper cavities were used to generate the radio frequency electric field that was used to accelerate electrons and positrons around the 27-km Large Electron-Positron (LEP) collider at CERN, which ran from 1989 to 2000. The copper cavities were gradually replaced from 1996 with new superconducting cavities allowing the collision energy to rise from 90 GeV to 200 GeV by mid-1999.

In oceans, estuaries, and rivers, nitrification is an important nitrate source, and stable isotopes of nitrate are often used to investigate recycling processes (e.g. remineralisation, nitrification) in the water column. Nitrification is a two-step process, where ammonia is oxidised via nitrite to nitrate. Nitrite usually does not accumulate in natural environments, which makes it difficult to study the single isotope effect of ammonia oxidation or nitrite oxidation in natural systems. However, during an exceptional flood in the Elbe River in June 2013, we found a unique co-occurrence of ammonium, nitrite, and nitrate in the water column, returning towards normal summer conditions within 1 week. Over the course of the flood, we analysed the evolution of δ15N-NH4+ and δ15N-NO2- in the Elbe River. In concert with changes in suspended particulate matter (SPM) and δ15N SPM, as well as nitrate concentration, δ15N-NO3- and δ18O-NO3-, we calculated apparent isotope effects during net nitrite and nitrate consumption. During the flood event, > 97 % of total reactive nitrogen was nitrate, which was leached from the catchment area and appeared to be subject to assimilation. Ammonium and nitrite concentrations increased to 3.4 and 4.4 µmol L-1, respectively, likely due to remineralisation, nitrification, and denitrification in the water column. δ15N-NH4+ values increased up to 12 ‰, and δ15N-NO2- ranged from -8.0 to -14.2 ‰. Based on this, we calculated an apparent isotope effect 15ɛ of -10.0 ± 0.1 ‰ during net nitrite consumption, as well as an isotope effect 15ɛ of -4.0 ± 0.1 ‰ and 18ɛ of -5.3 ± 0.1 ‰ during net nitrate consumption. On the basis of the observed nitrite isotope changes, we evaluated different nitrite uptake processes in a simple box model. We found that a regime of combined riparian denitrification and 22 to 36 % nitrification fits best with measured data for the nitrite concentration decrease and isotope increase.

This critical volume provides an in-depth presentation of copper wire bonding technologies, processes and equipment, along with the economic benefits and risks. Due to the increasing cost of materials used to make electronic components, the electronics industry has been rapidly moving from high cost gold to significantly lower cost copper as a wire bonding material. However, copper wire bonding has several process and reliability concerns due to its material properties. Copper Wire Bonding book lays out the challenges involved in replacing gold with copper as a wire bond material, and includes the bonding process changes—bond force, electric flame off, current and ultrasonic energy optimization, and bonding tools and equipment changes for first and second bond formation. In addition, the bond–pad metallurgies and the use of bare and palladium-coated copper wires on aluminum are presented, and gold, nickel and palladium surface finishes are discussed. The book also discusses best practices and re...

The effect of lead and copper on apical segments of Gracilaria domingensis was examined. Over a period of 7 days, the segments were cultivated with concentrations of 5 and 10 ppm under laboratory conditions. The samples were processed for light, confocal, and electron microscopy, as well as histochemistry, to evaluate growth rates, mitochondrial activity, protein levels, chlorophyll a, phycobiliproteins, and carotenoids. After 7 days of exposure to lead and copper, growth rates were slower than control, and biomass loss was observed on copper-treated plants. Ultrastructural damage was primarily observed in the internal organization of chloroplasts and cell wall thickness. X-ray microanalysis detected lead in the cell wall, while copper was detected in both the cytoplasm and cell wall. Moreover, lead and copper exposure led to photodamage of photosynthetic pigments and, consequently, changes in photosynthesis. However, protein content and glutathione reductase activity decreased only in the copper treatments. In both treatments, decreased mitochondrial NADH dehydrogenase activity was observed. Taken together, the present study demonstrates that (1) heavy metals such as lead and copper negatively affect various morphological, physiological, and biochemical processes in G. domingensis and (2) copper is more toxic than lead in G. domingensis.

To produce cellular energy, cyanobacteria reduce nitrate as the preferred pathway over proton reduction (H2 evolution) by catabolizing glycogen under dark anaerobic conditions. This competition lowers H2 production by consuming a large fraction of the reducing equivalents (NADPH and NADH). To eliminate this competition, we constructed a knockout mutant of nitrate reductase, encoded by narB, in Synechococcus sp. PCC 7002. As expected, ΔnarB was able to take up intracellular nitrate but was unable to reduce it to nitrite or ammonia, and was unable to grow photoautotrophically on nitrate. During photoautotrophic growth on urea, ΔnarB significantly redirects biomass accumulation into glycogen at the expense of protein accumulation. During subsequent dark fermentation, metabolite concentrations--both the adenylate cellular energy charge (∼ATP) and the redox poise (NAD(P)H/NAD(P))--were independent of nitrate availability in ΔnarB, in contrast to the wild type (WT) control. The ΔnarB strain diverted more reducing equivalents from glycogen catabolism into reduced products, mainly H2 and d-lactate, by 6-fold (2.8% yield) and 2-fold (82.3% yield), respectively, than WT. Continuous removal of H2 from the fermentation medium (milking) further boosted net H2 production by 7-fold in ΔnarB, at the expense of less excreted lactate, resulting in a 49-fold combined increase in the net H2 evolution rate during 2 days of fermentation compared to the WT. The absence of nitrate reductase eliminated the inductive effect of nitrate addition on rerouting carbohydrate catabolism from glycolysis to the oxidative pentose phosphate (OPP) pathway, indicating that intracellular redox poise and not nitrate itself acts as the control switch for carbon flux branching between pathways.

Full Text Available Proanthocyanidins (PAs contribute to poplar defense mechanisms against biotic and abiotic stresses. Transcripts of PA biosynthetic genes accumulated rapidly in response to infection by the fungus Marssonina brunnea f.sp. multigermtubi, treatments of salicylic acid (SA and wounding, resulting in PA accumulation in poplar leaves. Anthocyanidin reductase (ANR and leucoanthocyanidin reductase (LAR are two key enzymes of the PA biosynthesis that produce the main subunits: (+-catechin and (--epicatechin required for formation of PA polymers. In Populus, ANR and LAR are encoded by at least two and three highly related genes, respectively. In this study, we isolated and functionally characterized genes PtrANR1 and PtrLAR1 from P. trichocarpa. Phylogenetic analysis shows that Populus ANR1 and LAR1 occurr in two distinct phylogenetic lineages, but both genes have little difference in their tissue distribution, preferentially expressed in roots. Overexpression of PtrANR1 in poplar resulted in a significant increase in PA levels but no impact on catechin levels. Antisense down-regulation of PtrANR1 showed reduced PA accumulation in transgenic lines, but increased levels of anthocyanin content. Ectopic expression of PtrLAR1 in poplar positively regulated the biosynthesis of PAs, whereas the accumulation of anthocyanin and flavonol was significantly reduced (P<0.05 in all transgenic plants compared to the control plants. These results suggest that both PtrANR1 and PtrLAR1 contribute to PA biosynthesis in Populus.

Full Text Available Anthocyanidin reductase (ANR and leucoanthocyanidin reductase (LAR play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions.

Full Text Available BACKGROUND: Infusion of sodium nitrite could provide sustained therapeutic concentrations of nitric oxide (NO for the treatment of a variety of vascular disorders. The study was developed to determine the safety and feasibility of prolonged sodium nitrite infusion. METHODOLOGY: Healthy volunteers, aged 21 to 60 years old, were candidates for the study performed at the National Institutes of Health (NIH; protocol 05-N-0075 between July 2007 and August 2008. All subjects provided written consent to participate. Twelve subjects (5 males, 7 females; mean age, 38.8±9.2 years (range, 21-56 years were intravenously infused with increasing doses of sodium nitrite for 48 hours (starting dose at 4.2 µg/kg/hr; maximal dose of 533.8 µg/kg/hr. Clinical, physiologic and laboratory data before, during and after infusion were analyzed. FINDINGS: The maximal tolerated dose for intravenous infusion of sodium nitrite was 267 µg/kg/hr. Dose limiting toxicity occurred at 446 µg/kg/hr. Toxicity included a transient asymptomatic decrease of mean arterial blood pressure (more than 15 mmHg and/or an asymptomatic increase of methemoglobin level above 5%. Nitrite, nitrate, S-nitrosothiols concentrations in plasma and whole blood increased in all subjects and returned to preinfusion baseline values within 12 hours after cessation of the infusion. The mean half-life of nitrite estimated at maximal tolerated dose was 45.3 minutes for plasma and 51.4 minutes for whole blood. CONCLUSION: Sodium nitrite can be safely infused intravenously at defined concentrations for prolonged intervals. These results should be valuable for developing studies to investigate new NO treatment paradigms for a variety of clinical disorders, including cerebral vasospasm after subarachnoid hemorrhage, and ischemia of the heart, liver, kidney and brain, as well as organ transplants, blood-brain barrier modulation and pulmonary hypertension. CLINICAL TRIAL REGISTRATION INFORMATION: http

The NO reductase from Paracoccus denitrificans reduces NO to N2O (2NO + 2H(+) + 2e(-) → N2O + H2O) with electrons donated by periplasmic cytochrome c (cytochrome c-dependent NO reductase; cNOR). cNORs are members of the heme-copper oxidase superfamily of integral membrane proteins, comprising the O2-reducing, proton-pumping respiratory enzymes. In contrast, although NO reduction is as exergonic as O2 reduction, there are no protons pumped in cNOR, and in addition, protons needed for NO reduction are derived from the periplasmic solution (no contribution to the electrochemical gradient is made). cNOR thus only needs to transport protons from the periplasm into the active site without the requirement to control the timing of opening and closing (gating) of proton pathways as is needed in a proton pump. Based on the crystal structure of a closely related cNOR and molecular dynamics simulations, several proton transfer pathways were suggested, and in principle, these could all be functional. In this work, we show that residues in one of the suggested pathways (denoted pathway 1) are sensitive to site-directed mutation, whereas residues in the other proposed pathways (pathways 2 and 3) could be exchanged without severe effects on turnover activity with either NO or O2. We further show that electron transfer during single-turnover reduction of O2 is limited by proton transfer and can thus be used to study alterations in proton transfer rates. The exchange of residues along pathway 1 showed specific slowing of this proton-coupled electron transfer as well as changes in its pH dependence. Our results indicate that only pathway 1 is used to transfer protons in cNOR.

Background: Sodium or potassium nitrite is widely used as a curing agent in sausages and other cured meat products. Nitrite has strong antimicrobial and antioxidant effects and generates cured meat color. Nitrite, however, can react with secondary or tertiary amines in meat to form carcinogenic, teratogenic and mutagenic N-nitroso compounds. Several findings have been suggested that high consumption of processed meat may increase the risk of cancer, and emphasized that dietary nitrosamines ar...

The responses of a large suite of biochemical and genetic parameters were evaluated in tissues (liver, gills, muscle and erythrocytes) of the estuarine guppy Poecilia vivipara exposed to waterborne copper in salt water (salinity 24 ppt). Activities of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase, and glutathione S-transferase), metallothionein-like protein concentration, reactive oxygen species (ROS) content, antioxidant capacity against peroxyl radicals (ACAP), and lipid peroxidation (LPO) were evaluated in liver, gills, and muscle. Comet assay score and nuclear abnormalities and micronucleated cell frequency were analyzed in peripheral erythrocytes. The responses of these parameters were evaluated in fish exposed (96 h) to environmentally relevant copper concentrations (5, 9 and 20 μg L⁻¹). In control and copper-exposed fish, no mortality was observed over the experimental period. Almost all biochemical and genetic parameters proved to be affected by waterborne copper exposure. However, the response of catalase activity in liver, ROS, ACAP and LPO in muscle, gills and liver, and DNA damages in erythrocytes clearly showed to be dependent on copper concentration in salt water. Therefore, the use of these parameters could be of relevance in the scope of biomonitoring programs in salt water environments contaminated with copper.

Full Text Available Nitrite poisoning in pigs was suspected when 4 of 18 pigs died in a piggery near Ellisras in the Northern Province. The pigs showed typical brownish discolouration of the blood at autopsy. It was established that they ingested vegetable tops and weeds from the adjacent garden as part of their daily ration. Of the available plants, only Capsella bursa-pastoris contained nitrites. The drinking water and some of the other plants tested positive for nitrates but not for nitrites. This is the first report of suspected nitrite poisoning in pigs caused by Capsella bursa-pastoris.

It has been reported that mitochondrial aldehyde dehydrogenase (ALDH2) catalyzes the formation of glyceryl dinitrate and inorganic nitrite from glyceryl trinitrate (GTN), leading to an increase in cGMP and vasodilation in the coronary and systemic vascular beds. However, the role of nitric oxide (NO) formed from nitrite in mediating the response to GTN in the pulmonary vascular bed is uncertain. The purpose of the present study was to determine if nitrite plays a role in mediating vasodilator responses to GTN. In this study, intravenous injections of GTN and sodium nitrite decreased pulmonary and systemic arterial pressures and increased cardiac output. The decreases in pulmonary arterial pressure under baseline and elevated tone conditions and decreases in systemic arterial pressure in response to GTN and sodium nitrite were attenuated by cyanamide, an ALDH2 inhibitor, whereas responses to the NO donor, sodium nitroprusside (SNP), were not altered. The decreases in pulmonary and systemic arterial pressure in response to GTN and SNP were not altered by allopurinol, an inhibitor of xanthine oxidoreductase, whereas responses to sodium nitrite were attenuated. GTN was approximately 1,000-fold more potent than sodium nitrite in decreasing pulmonary and systemic arterial pressures. These results suggest that ALDH2 plays an important role in the bioactivation of GTN and nitrite in the pulmonary and systemic vascular beds and that the reduction of nitrite to vasoactive NO does not play an important role in mediating vasodilator responses to GTN in the intact chest rat.

To evaluate the relationship between atmospheric nitrogen dioxide exposure and the development of allergic diseases, the effects of nitrite as a chemical product of inhaled nitrogen dioxide on mast cell functions were investigated. We have studied nitride-induced histamine release from two functionally distinct mast cell populations, namely peritoneal mast cells (PMC) and intestinal mucosal mast cells (IMMC) of Nippostrongylus brasiliensis-infected rats. High concentrations of nitrite alone (10, 20, and 50 mM) induced histamine release from IMMC, but not from PMC. Moreover, histamine release from PMC and IMMC stimulated with sensitizing antigen was significantly enhanced by pretreatment with 50 mM nitrite or nitrate. No differences in histamine release from nitrite-treated and control PMC were seen below 1 mM. To investigate the effect of nitrite on tumor cell cytotoxic activity, PMC were incubated with various concentrations of nitrite. Pretreatment with 5 and 50 mM nitrite markedly depressed tumor necrosis factor (TNF)-[alpha]-dependent natural cytotoxicity of PMC for the tumor target WEHI-164. Thus, high concentrations of nitrite enhanced mast cell histamine release, but depressed TNF-[alpha]-dependent cytotoxicity. However, low concentrations of nitrite (<1 mM) that would normally be produced by short-term atmospheric exposure to nitrogen dioxide may have no significant effects on mast cell functions. 27 refs., 3 figs., 1 tab.

Cobalt(III) 5, 10, 15-tris(4-tert-butylphenyl) corrole was synthesized and incorporated into plasticized poly(vinyl chloride) membranes and studied as a neutral carrier ionophore via potentiometry. This cobalt(III) complex has binding affinity to nitrite, and the resulting membrane electrode yields reversible and Nernstian response toward nitrite. Enhanced nitrite selectivity is observed over other anions, including lipophilic anions such as thiocyanate and perchlorate when an appropriate amount of lipophilic cationic sites are added to the membrane phase. Detection limit to nitrite is ca. 5 µM. Using tributylphosphate as the plasticizer with the cobalt(III) corrole species yields electrodes with enhanced nitrate selectivity.

Copper is a transition metal that has been linked to pathological and beneficial effects in neurodegenerative diseases. In Parkinson's disease, free copper is related to increased oxidative stress, alpha-synuclein oligomerization, and Lewy body formation. Decreased copper along with increased iron has been found in substantia nigra and caudate nucleus of Parkinson's disease patients. Copper influences iron content in the brain through ferroxidase ceruloplasmin activity; therefore decreased pr...

There are currently no FDA-approved antidotes for H2S/sulfide intoxication. Sodium nitrite, if given prophylactically to Swiss Webster mice, was shown to be highly protective against the acute toxic effects of sodium hydrosulfide (∼LD40 dose) with both agents administered by intraperitoneal injections. However, sodium nitrite administered after the toxicant dose did not detectably ameliorate sulfide toxicity in this fast-delivery, single-shot experimental paradigm. Nitrite anion was shown to rapidly produce NO in the bloodstream, as judged by the appearance of EPR signals attributable to nitrosylhemoglobin and methemoglobin, together amounting to less than 5% of the total hemoglobin present. Sulfide-intoxicated mice were neither helped by the supplemental administration of 100% oxygen nor were there any detrimental effects. Compared to cyanide-intoxicated mice, animals surviving sulfide intoxication exhibited very short knockdown times (if any) and full recovery was extremely fast (∼15 min) irrespective of whether sodium nitrite was administered. Behavioral experiments testing the ability of mice to maintain balance on a rotating cylinder showed no motor impairment up to 24 h post sulfide exposure. It is argued that antagonism of sulfide inhibition of cytochrome c oxidase by NO is the crucial antidotal activity of nitrite rather than formation of methemoglobin.

A novel fiber optic spectrophotometric method for nitrite determination in different samples is suggested, based on the reaction of nitrite with Safranin O in acidic medium to form a diazo-safranin, which is subsequently coupled with pyrogallol in alkaline medium to form a highly stable, red azo dye, followed by cloud point extraction (CPE) using a mixed micelle of a nonionic surfactant, Triton X-114, with an anionic surfactant, sodium dodecyl sulphate (SDS). The reaction and extraction conditions (e.g., acidity for diazotization and alkalinity for pyrogallol coupling, and other reagent concentrations, time, and tolerance to other ions) were optimized. Linearity was obeyed in a concentration range up to 230 μg L(-1), and the detection limit of the method is 0.5 μg L(-1) of nitrite ion. The molar absorptivity for nitrite of the Safranin-diazonium salt (ɛ(610 nm) =4 × 10(3) L mol(-1)cm(-1)) existing in literature was greatly enhanced by pyrogallol coupling and CPE enrichment (ɛ(592 nm)=1.39 × 10(5) L mol(-1)cm(-1)). The method was applied to the determination of nitrite in tap water, lake water and milk samples with an optimal preconcentration factor of 20.

Nitrate and nitrite ions are of considerable interest, both for their widespread use in commercial and research contexts and because of their central role in the global nitrogen cycle. The chemistry of atmospheric aerosols, wherein nitrate is abundant, has been found to depend on the interfacial behavior of ionic species. The interfacial behavior of ions is determined largely by their hydration properties; consequently, the study of the hydration and interfacial behavior of nitrate and nitrite comprises a significant field of study. In this work, we describe the study of aqueous solutions of sodium nitrate and nitrite via X-ray absorption spectroscopy (XAS), interpreted in light of first-principles density functional theory electronic structure calculations. Experimental and calculated spectra of the nitrogen K-edge XA spectra of bulk solutions exhibit a large 3.7 eV shift between the XA spectra of nitrate and nitrite resulting from greater stabilization of the nitrogen 1s energy level in nitrate. A similar shift is not observed in the oxygen K-edge XA spectra of NO{sub 3}{sup −} and NO{sub 2}{sup −}. The hydration properties of nitrate and nitrite are found to be similar, with both anions exhibiting a similar propensity towards ion pairing.

An efficient luminescence energy transfer (LET) system based on terbium(III)-sodium hexametaphosphate (Tb/SHMP) chelates as donor and 4-((4-(2-aminoethylamino)naphthalen-1-yl)diazenyl)benzenesulfonic acid dihydrochloride (ANDBS) as acceptor was developed for sensitive determination of trace nitrite. Stable and strong fluorescence Tb/SHMP chelates were prepared in aqueous solution. Based on Griess Reaction, ANDBS was generated by the quantitative reaction of nitrite, sulfanilamide and N-(1-naphtyl)-ethylenediamine dihydrochloride (N1NED). The degree of the overlap was effective for LET between the emission spectrum of Tb/SHMP chelates and the absorption spectrum of ANDBS. Based on the luminescence intensity quenching of Tb/SHMP chelates in proportion to the trace amounts of nitrite, a new assay for the selective and sensitive determination of nitrite was developed. Under the optimum conditions, the linear calibration graph was obtained with a linear range of 0.00040-0.20 {mu}g mL{sup -1} (R=0.99657). The detection limit of NO{sub 2}{sup -} was 0.00010 {mu}g mL{sup -1} (R=0.99657). The method was applied successfully to the determination of nitrite for synthetic samples.

Nitrite, at an environmentally relevant concentration, was significantly reduced with iron (hydr)oxides mediated by Shewanella oneidensis MR-1. The average nitrite removal rates of 1.28±0.08 and 0.65±0.02(mgL(-1))h(-1) were achieved with ferrihydrite and magnetite, respectively. The results showed that nitrite removal was able to undergo multiple redox cycles with iron (hydr)oxides mediated by Shewanella oneidensis MR-1. During the bioreduction of the following cycles, biogenic Fe(II) was subsequently chemically oxidized to Fe(III), which is associated with nitrite reduction. There was 11.18±1.26mgL(-1) of NH4(+)-N generated in the process of redox cycling of ferrihydrite. Additionally, results obtained by using X-ray diffraction showed that ferrihydrite and magnetite remained mainly stable in the system. This study indicated that redox cycling of Fe in iron (hydr)oxides was a potential process associated with NO2(-)-N removal from solution, and reduced most nitrite abiotically to gaseous nitrogen species.

A simple and sensitive spectrofluorimetric method was developed for the determination of nitrite in environmental and food samples. The method was based on the selective reaction of o-phenylenediamine with nitrite in acidic medium to form benzotriazole, which exhibited strong fluorescence at 568 nm with excitation at 420 nm in alkaline medium. The detection limit and sensitivity of the proposed method were improved by hydroxypropyl-β-cyclodextrin through complexation. The linear calibration range for nitrite was 0.04-0.8 μg mL-1 with a detection limit of 13.6 ng mL-1 (S/N = 3.29). The relative standard deviation for ten determinations of 0.1 and 0.4 μg mL-1 nitrite were 1.38% and 2.01%, respectively. Twenty-eight coexistent ions were examined, and no serious interference for most of ions was observed. The proposed method was successfully applied for the determination of nitrite in the water, sausage and soil samples with recoveries of 95.5-108.5%. The results were in good agreement with the recommended AOAC method.

An anaerobic-oxic(A/O)biological phosphorus removal reactor was operated to study the effect of nitrite on phosphate uptake.The phosphorus uptake profile was determined under different operating conditions.The results indicated that in addition to oxygen and nitrate(DPBNa,nitrate denitrifying phosphorus removal),to some extent,nitrite could also serve as an electron acceptor to achieve nitrite denitrifying phosphorus removal(DPBNi).The quantity and rate of phosphorus uptake of DPBNi,however,were evidently lower than that of DPBNa.The experiment results revealed that nitrite would bring toxic action to phosphate accumulating organisms(PAOs)when NO2- -N≥93]7 mg/L.The nitrite existing in the anoxic reactor made no difference to the quantity and rate of denitrifying phosphorus removal,but it could reduce the consumption of nitrate.Moreover,the data showed that the aerobic phosphate uptake of DPBNi was lower than that of anaerobic phosphorus-released sludge in a traditional A/O process.However,there was not much difference between these two kinds of sludge in terms of the total phosphorus uptake quantity and the effluent quality.

This study aims to understand the mechanisms of nitrite appearance during wastewater denitrification by biofilters, focusing on the role of the carbon source. Experiments were carried out at lab-scale (batch tests) and full-scale plant (Parisian plant, capacities of 240,000 m(3) day(-1)). Results showed that the nature of the carbon source affects nitrite accumulation rates. This accumulation is low, 0.05 to 0.10 g N-NO2(-) per g N-NO3(-) eliminated, for alcohols such as methanol, ethanol, or glycerol. The utilization of glycerol leads to fungal development causing clogging of the biofilters. This fungal growth and consequent clogging exclude this carbon source, with little nitrite accumulation, as carbon source for denitrification. Whatever the carbon source, the C/N ratio in the biofilter plays a major role in the appearance of residual nitrite; an optimal C/N ratio from 3.0 to 3.2 allows a complete denitrification without any nitrite accumulation.

It remains challenging to integrate nitritation and anammox in ecologically engineered treatment systems such as passive biofilters that are packed with natural materials and have low energy inputs. This study explored the factors influencing nitritation-anammox through parallel operation of two laboratory-scale biofilters packed with large and small marble chips respectively. Clean marble chips (mainly CaCO3) had an alkalinity dissolution rate of 130 mg CaCO3/kg marble d when water pH approached 6.5. Marble chips effectively increased water pH and provided sufficient alkalinity to support nitritation-anammox in the biofilters. Ammonium and total nitrogen removal decreased by 47 and 26%, respectively, when nutrients were not amended to influent. An influent nitrite concentration above 8.9 mg N/L could inhibit anammox in thin biofilms of biofilters. Nitritation-anammox was enhanced with a hydraulic retention time of 2 d relative to 7 d, likely due to enhanced air entrainment. Size of marble chips rarely made a significant difference in nitrogen removal, possibly due to sufficient surface area available for bacterial attachment and alkalinity dissolution.

Adverse effects associated with recreational inhalation of nitrites are usually mild and rarely life-threatening. We report a rare case of near-fatal methemoglobinemia after inhalation of amyl nitrite after aerosolizing the liquid using a compressed gas blower designed to clean photographic equipment that employed hydrofluoroalkane-134a as a propellant. A 31-year-old previously healthy male became dyspneic and fainted soon after the recreational inhalation of amyl nitrite aerosolized using a compressed gas blower. He was brought to the emergency department with severe cyanotic appearance and profound shock. Oxygen saturation was 82%, unresponsive to oxygen supply. His methemoglobin blood level was 52.2%. After 100 mg of methylene blue (2 mg/kg body weight) was administered intravenously, he recovered consciousness, and dyspnea and cyanosis subsided gradually. This case illustrates the extraordinary hazard of the use of a compressed gas blower in the recreational inhalation of nitrites. Prompt recognition and rapid antidotal treatment may adequately correct near-fatal overdose associated with recreational use of amyl nitrite.

This study presents a biological system combined upflow anaerobic sludge bed (UASB) with sequencing batch reactor (SBR) to treat ammonium-rich landfill leachate. The start-up and operation of the nitritation at low temperatures were investigated. The synergetic interaction of free ammonia (FA) inhibition on nitrite-oxidizing bacteria (NOB) and process control was used to achieve nitritation in the SBR. It is demonstrated that nitritation was successful y started up in the SBR at low temperatures (14.0 °C–18.2 °C) by using FA inhibition coupled with process control, and then was maintained for 482 days at normal/low temperature. Although ammonia-oxidizing bacteria (AOB) and NOB co-existed within bacterial clusters in the SBR sludge, AOB were confirmed to be dominant nitrifying population species by scanning electron microscopic (SEM) observation and fluorescence in situ hybridization (FISH) analysis. This confirmation not only emphasized that cultivating the appropriate bacteria is essential for achieving stable nitritation performance, but it also revealed that NOB activity was strongly inhibited by FA rather than being eliminated altogether from the system.

Full Text Available A selective and rapid spectrophotometric method for the determination of nitrite is presented. It is based on the reaction of nitrite with p-nitroaniline in acid medium to form diazonium ion, which is coupled with ethoxyethylenemaleic ester or ethylcyanoacetate in basic medium to form azo dyes, showing absorption maxima at 439 and 465 nm respectively. The method obeys Beer's law in the concentration range of 0.5-16 µg mL-1 of nitrite with ethoxyethylenemaleic ester and 0.2-18 µg mL-1 of nitrite with ethylcyanoacetate. The molar absorptivity and Sandell's sensitivity of p-nitroaniline-ethoxyethylenemaleic ester and p-nitroaniline-ethylcyanoacetate azo dyes are 5.04 X 10(4 L mol-1cm-1, 0.98 X 10-2 µg cm-2 and 1.21 X 10(4 L mol-1 cm-1, 0.98 X 10-2 µg cm-2 respectively. The optimum reaction conditions and other analytical parameters were evaluated. The method was successfully applied to the determination of nitrite in various water samples and soil samples.

Evidence from both mammalian and nonmammalian vertebrates indicates that intracardiac nitric oxide (NO) facilitates myocardial relaxation, ventricular diastolic distensibility, and, consequently, the Frank-Starling response, i.e., the preload-induced increase of cardiac output. Since nitrite ion (NO(2)(-)), the major storage pool of bioactive NO, recently emerged as a cardioprotective endogenous modulator, we explored its influence on the Frank-Starling response in eel, frog, and rat hearts, used as paradigms of fish, amphibians, and mammals, respectively. We demonstrated that, like NO, exogenous nitrite improves the Frank-Starling response in all species, as indicated by an increase of stroke volume and stroke work (eel and frog) and of left ventricular (LV) pressure and LVdP/dt max (rat), used as indexes of inotropism. Unlike in frog and rat, in eel, the positive influence of nitrite appeared to be dependent on NO synthase inhibition. In all species, the effect was sensitive to NO scavengers, independent on nitroxyl anion, and mediated by a cGMP/PKG-dependent pathway. Moreover, the nitrite treatment increased S-nitrosylation of lower-molecular-weight proteins in cytosolic and membrane fractions. These results suggest that nitrite acts as a physiological source of NO, modulating through different species-specific mechanisms, the stretch-induced intrinsic regulation of the vertebrate heart.

According to news published on March 30th, China’s largest copper producer--Jiangxi Copper alleged in its 2010 Financial Report Statement that it plans to improve its output of refined copper by 4.4% in 2011, to increase from 900,000 tonnes last year to 940,000 tons.

Research highlights: {yields} Two monomeric 90 kDa nitrate reductase active proteins from Shewanella gelidimarina. {yields} Sequence of napA from napEDABC-type operon and napA from NapDAGHB-type operon. {yields} Isolation of NAP as NapA or NapAB correlated with NapA P47E amino acid substitution. -- Abstract: The reduction of nitrate to nitrite in the bacterial periplasm occurs in the 90 kDa NapA subunit of the periplasmic nitrate reductase (NAP) system. Most Shewanella genomes contain two nap operons: napEDABC and napDAGHB, which is an unusual feature of this genus. Two native, monomeric, 90 kDa nitrate reductase active proteins were resolved by hydrophobic interaction chromatography from aerobic cultures of Shewanella gelidimarina replete with reduced nitrogen compounds. The 90 kDa protein obtained in higher yield was characterized as NapA by electronic absorption and electron paramagnetic resonance spectroscopies and was identified by LC/MS/MS and MALDI-TOF/TOF MS as NapA from the napEDABC-type operon. The other 90 kDa protein, which was unstable and produced in low yields, was posited as NapA from the napDAGHB-type operon. Two napA genes have been sequenced from the napEDABC-type and napDAGHB-type operons of S. gelidimarina. Native NAP from S. putrefaciens was resolved as one NapA monomer and one NapAB heterodimer. Two amino acid substitutions in NapA correlated with the isolation of NAP as a NapA monomer or a NapAB heterodimer. The resolution of native, redox-active NapA isoforms in Shewanella provides new insight into the respiratory versatility of this genus, which has implications in bioremediation and the assembly of microbial fuel cells.

Copper is one of essential trace elements. Copper deficiency lead to growth and developmental failure and/or neurological dysfunction. However, excess copper is also problems for human life. There are two disorders of inborn error of copper metabolism, Menkes disease and Wilson disease. Menkes disease is an X linked recessive disorder with copper deficiency and Wilson disease is an autosomal recessive disorder with copper accumulation. These both disorders result from the defective functioning of copper transport P-type ATPase, ATP7A of Menkes disease and ATP7B of Wilson disease. In this paper, the author describes about copper metabolism of human, and clinical feature, diagnosis and treatment of Menkes disease and Wilson disease.

Simpson, Philippa J.L. [School of Chemistry, University of Sydney, New South Wales 2006 (Australia); Codd, Rachel, E-mail: rachel.codd@sydney.edu.au [School of Chemistry, University of Sydney, New South Wales 2006 (Australia); School of Medical Sciences (Pharmacology) and Bosch Institute, University of New South Wales, New South Wales 2006 (Australia)

2011-11-04

Highlights: Black-Right-Pointing-Pointer Cold-adapted phenotype of NapA from the Antarctic bacterium Shewanella gelidimarina. Black-Right-Pointing-Pointer Protein homology model of NapA from S. gelidimarina and mesophilic homologue. Black-Right-Pointing-Pointer Six amino acid residues identified as lead candidates governing NapA cold adaptation. Black-Right-Pointing-Pointer Molecular-level understanding of designing cool-temperature in situ oxyanion sensors. -- Abstract: The reduction of nitrate to nitrite is catalysed in bacteria by periplasmic nitrate reductase (Nap) which describes a system of variable protein subunits encoded by the nap operon. Nitrate reduction occurs in the NapA subunit, which contains a bis-molybdopterin guanine dinucleotide (Mo-MGD) cofactor and one [4Fe-4S] iron-sulfur cluster. The activity of periplasmic nitrate reductase (Nap) isolated as native protein from the cold-adapted (psychrophilic) Antarctic bacterium Shewanella gelidimarina (Nap{sub Sgel}) and middle-temperature adapted (mesophilic) Shewanella putrefaciens (Nap{sub Sput}) was examined at varied temperature. Irreversible deactivation of Nap{sub Sgel} and Nap{sub Sput} occurred at 54.5 and 65 Degree-Sign C, respectively. When Nap{sub Sgel} was preincubated at 21-70 Degree-Sign C for 30 min, the room-temperature nitrate reductase activity was maximal and invariant between 21 and 54 Degree-Sign C, which suggested that Nap{sub Sgel} was poised for optimal catalysis at modest temperatures and, unlike Nap{sub Sput}, did not benefit from thermally-induced refolding. At 20 Degree-Sign C, Nap{sub Sgel} reduced selenate at 16% of the rate of nitrate reduction. Nap{sub Sput} did not reduce selenate. Sequence alignment showed 46 amino acid residue substitutions in Nap{sub Sgel} that were conserved in NapA from mesophilic Shewanella, Rhodobacter and Escherichia species and could be associated with the Nap{sub Sgel} cold-adapted phenotype. Protein homology modeling of Nap{sub Sgel} using a

Reactive Oxygen Species (ROS) are involved in plant biomass degradation by fungi and development of fungal structures. While the ROS-generating NADPH oxidases from filamentous fungi are under strong scrutiny, much less is known about the related integral Membrane (or Ferric) Reductases (IMRs). Here, we present a survey of these enzymes in 29 fungal genomes covering the entire available range of fungal diversity. IMRs are present in all fungal genomes. They can be classified into at least 24 families, underscoring the high diversity of these enzymes. Some are differentially regulated during colony or fruiting body development, as well as by the nature of the carbon source of the growth medium. Importantly, functional characterization of IMRs has been made on proteins belonging to only two families, while nothing or very little is known about the proteins of the other 22 families.

The proanthocyanidins (PAs), a subgroup of flavonoids, accumulate to levels of approximately 10% total dry weight of cacao seeds. PAs have been associated with human health benefits and also play important roles in pest and disease defense throughout the plant. To dissect the genetic basis of PA biosynthetic pathway in cacao (Theobroma cacao), we have isolated three genes encoding key PA synthesis enzymes, anthocyanidin synthase (ANS), anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR). We measured the expression levels of TcANR, TcANS and TcLAR and PA content in cacao leaves, flowers, pod exocarp and seeds. In all tissues examined, all three genes were abundantly expressed and well correlated with PA accumulation levels, suggesting their active roles in PA synthesis. Overexpression of TcANR in an Arabidopsis ban mutant complemented the PA deficient phenotype in seeds and resulted in reduced anthocyanidin levels in hypocotyls. Overexpression of TcANS in tobacco resulted in increased content of both anthocyanidins and PAs in flower petals. Overexpression of TcANS in an Arabidopsis ldox mutant complemented its PA deficient phenotype in seeds. Recombinant TcLAR protein converted leucoanthocyanidin to catechin in vitro. Transgenic tobacco overexpressing TcLAR had decreased amounts of anthocyanidins and increased PAs. Overexpressing TcLAR in Arabidopsis ldox mutant also resulted in elevated synthesis of not only catechin but also epicatechin. Our results confirm the in vivo function of cacao ANS and ANR predicted based on sequence homology to previously characterized enzymes from other species. In addition, our results provide a clear functional analysis of a LAR gene in vivo.

Most high Mr thioredoxin reductases (TRs) have the unusual feature of utilizing a vicinal disulfide bond (Cys1-Cys2) which form an eight-membered ring during the catalytic cycle. Many eukaryotic TRs have replaced the Cys2 position of the dyad with the rare amino acid selenocysteine (Sec). Here we demonstrate that Cys- and Sec-containing TRs are distinguished by the importance each class of enzymes places on the 8-membered ring structure in the catalytic cycle. This hypothesis was explored by studying the truncated enzyme missing the C-terminal ring structure in conjunction with oxidized peptide substrates to investigate the reduction and opening of this dyad. The peptide substrates were identical in sequence to the missing part of the enzyme, containing either a disulfide or selenylsulfide linkage, but were differentiated by the presence (cyclic) and absence (acyclic) of the ring structure. The ratio of these turnover rates informs that the ring is only of modest importance for the truncated mouse mitochondrial Sec-TR (ring/no ring = 32), while the ring structure is highly important for the truncated Cys-TRs from D. melanogaster and C. elegans (ring/no ring > 1000). All three enzymes exhibit a similar dependence upon leaving group pKa as shown by the use of the acyclic peptides as substrates. These two factors can be reconciled for Cys-TRs if the ring functions to simultaneously allow for attack by a nearby thiolate while correctly positioning the leaving group sulfur atom to accept a proton from the enzymic general acid. For Sec-TRs the ring is unimportant because the lower pKa of the selenol relative to a thiol obviates its need to be protonated upon S-Se bond scission and permits physical separation of the selenol and the general acid. Further study of the biochemical properties of the truncated Cys and Sec TR enzymes demonstrates that the chemical advantage conferred on the eukaryotic enzyme by a selenol is the ability to function at acidic pH. PMID:18986163

Twenty-four 4-dimethylaminoazobenzenes (DABs) in which systematic structural modifications have been made in the prime ring have been studied for substrate specificity for microsomal azo reductase. The DABs were also evaluated for carcinogenicity and it was found that there was no correlation between carcinogenicity and extent of azo bond cleavage by azo reductase. While any substituent in the prime ring reduces the rate of cleavage of the azo bond relative to the unsubstituted dye, there is a correlation between substituent size and susceptibility to the enzyme. Substituent size was also found to be a significant factor in the induction of hepatomas by the dyes. Preliminary studies have shown that there appears to be a positive correlation between microsomal riboflavin content and the activity of the azo reductase.

Full Text Available Background: Because of high consumption of nitrite in processed (fast foods and high level of nitrite in water, soil and ecosystem, nitrite can endanger humans health. In this study the effects of sodium nitrite on aorta was examined in adult male rats. Materials and Methods : In the present study, 30 Wistar adult male rats were randomly divided into three groups of 10, including; control group. First experimental group that received low dose of sodium nitrite (175 mg/kg.bw, second experimental group that received high dose of sodium nitrite (350 mg/kg.bw. They were examined for 60 days. The rats got sodium nitrite through drinking water. At the end of the experiment the rats were taken to the anesthesia jar and based on ether principles, they anesthetized with ether and their blood samples were collected from their hearts. Then their aorta were extracted from their bodies and the tissue sections were prepared for testing tissue changes. Features such as histological features of aorta (morphometric and morphologic features were analyzed. The samples were stained with masson trichrome and Hematoxylin- Eosin methods. The internal media layer was measured with Image tool software. Then the amount of nitrite oxide in their blood were tested. At the end results were analyzed by 17 version of SPSS software and ANOVA test was run. Results: The results of this study showed that thickness of medial layer in two experimental group that received low and high dose of sodium nitrite compared with the control group decreased (p 0.05, and the group that received of high dose of sodium nitrite showed irregular and non- uniform state in aortic media layer. Conclusion: The finding of this study indicated that consumption of sodium nitrite in long term can induce damage in artries tissue.

To investigate the role of nitric oxide (NO) in steroid-induced femoral head necrosis, NO production was measured indirectly as nitrite in serum and synovial fluid samples from patients with steroid-induced femoral head necrosis together with serum samples from healthy volunteers matched for age and sex. The results showed that:① serum nitrite concentration in patients with femoral head necrosis were significantly lower than that in controls (P＜0.001); ② nitrite level of synovial fluid was markedly higher than that of serum. In addition, there was a positive correlation between them (r＝0.378,P＜0.05). We reach the conclusion:① NO is synthesized by synovium and chondrocytes; ② the decreased NO concentration in serum suggests a protective role in steroid-induced femoral head necrosis.

AIM: To investigate the diagnostic efficacy of leukocyte esterase and nitrite reagent strips for bedside diagnosis of spontaneous bacterial peritonitis (SBP).METHODS: A total of 63 consecutive patients with cirrhotic ascites (38 male, 25 female) tested between April 2005 and July 2006 were included in the study. Bedside reagent strip testing was performed on ascitic fluid and the results compared to manual cell counting and ascitic fluid culture. SBP was defined as having a olymorphonuclear ascites count of ≥ 250/mm3.RESULTS: Fifteen samples showed SBP. The sensitivity,specificity, positive and negative predictive values of the leukocyte esterase reagent strips were; 93%, 100%, 100%, and 98%, respectively. The sensitivity,specificity, positive and negative predictive value of the nitrite reagent strips were 13%, 93%, 40%, and 77%, respectively. The combination of leukocyte esterase and nitrite reagents strips did not yield statistically significant effects on diagnostic accuracy. CONCLUSION: Leukocyte esterase reagent strips may provide a rapid, bedside diagnostic test for SBP.

We evaluated the effect of hypertension on hemodynamic responses and serum nitrite concentrations in normotensive (NT) and deoxycorticosteron acetate (DOCA)-Salt hypertensive (HT) rats. Uncontrolled hemorrhagic shock was induced in NT and HT rats (n=7 each) by preliminary bleed of 25 ml/kg followed by a 75% tail amputation. The mean arterial pressure (MAP), heart rate and serum nitrite were measured pre-hemorrhage and during hemorrhage. Changes in time-averaged MAP after hemorrhage were significantly greater in HT group than NT. After resuscitation, the HT rats failed to restore MAP to baseline level. Serum nitrite level in both groups was significantly increased during shock period. Survival rate of HT animals was lower than NT group, although it was not statistically significant. Marked reduction of MAP and less improvement after resuscitation suggested the less adaptation of cardiovascular system in HT animals which may interfere with management of these subjects during uncontrolled hemorrhagic shock.

Nitrite has been postulated to provide a reservoir for conversion to nitric oxide (NO), especially in tissues with reduced oxygen levels as in the fetus. Nitrite would thus provide local vasodilatation and restore a balance between oxygen supply and need, a putative mechanism of importance especially in the brain. The current experiments test the hypothesis that exogenous nitrite acts as a vasodilator in the cephalic vasculature of the intact, near term fetal sheep. Fetuses were first instrumented to measure arterial blood pressure and carotid artery blood flow and then studied 4-5 days later while in utero without anaesthesia. Initially l-nitro-arginine (LNNA) was given to block endogenous NO production. Carotid resistance to flow increased 2-fold from 0.54 ± 0.01 (SEM) to 1.20 ± 0.08 mmHg min ml(-1) (in 13 fetuses, P fetal sheep.

A simple kinetic model was developed for describing nitrite oxidation by autotrophic aerobic nitrifiers in a continuous stirred tank reactor (CSTR), in which mixed (suspended and attached) growth conditions prevail. The CSTR system was operated under conditions of constant nitrite feed concentration and varying volumetric flow rates. Experimental data from steady-state conditions in the CSTR system and from batch experiments were used for the determination of the model's kinetic parameters. Model predictions were verified against experimental data obtained under transient operating conditions, when volumetric flow rate and nitrite feed concentration disturbances were imposed on the CSTR. The presented kinetic modeling procedure is quite simple and general and therefore can also be applied to other mixed growth biological systems.

Full Text Available Changes in day length enhance or suppress component of immune function in individuals of several species. The purpose of the present experiment was to study the role of photoperiodic manipulation on the nitric oxide production by splenic macrophages in the fresh-water snake, Natrix piscator. To study effect of photoperiod, animals were subjected to 24 hour continuous light and continuous dark for 30 days. Animals kept in natural day length served as control. At termination of experiments, animals were sacrificed, and spleen was excised. Macrophages were incubated for 24 hours and nitric oxide production was measured by measuring the nitrite concentration. Nitrite production was significantly decreased to the cultures obtained from the animals kept under continuous light. No change in nitrite concentration was found in animals kept under continuous dark, when compared to the animals kept under natural day length. The possible role of decreased melatonin synthesis in light is suggested to decrease the nitric oxide production.

Alkyl hydroperoxide reductase (AhpC) is known to detoxify peroxides and reactive sulfur species (RSS). However, the relationship between its expression and combating of abiotic stresses is still not clear. To investigate this relationship, the genes encoding the alkyl hydroperoxide reductase (ahpC) from Anabaena sp. PCC 7120 were introduced into E. coli using pGEX-5X-2 vector and their possible functions against heat, salt, carbofuron, cadmium, copper and UV-B were analyzed. The transformed E. coli cells registered significantly increase in growth than the control cells under temperature (47 {sup o}C), NaCl (6% w/v), carbofuron (0.025 mg ml{sup -1}), CdCl{sub 2} (4 mM), CuCl{sub 2} (1 mM), and UV-B (10 min) exposure. Enhanced expression of ahpC gene as measured by semi-quantitative RT-PCR under aforementioned stresses at different time points demonstrated its role in offering tolerance against multiple abiotic stresses.

Full Text Available Mitochondrial thioredoxin-glutathione reductase was purified from larval Taenia crassiceps (cysticerci. The preparation showed NADPH-dependent reductase activity with either thioredoxin or GSSG, and was able to perform thiol/disulfide exchange reactions. At 25∘C specific activities were 437 ± 27 mU mg-1 and 840 ± 49 mU mg-1 with thioredoxin and GSSG, respectively. Apparent Km values were 0.87 ± 0.04 μM, 41 ± 6 μM and 19 ± 10 μM for thioredoxin, GSSG and NADPH, respectively. Thioredoxin from eukaryotic sources was accepted as substrate. The enzyme reduced H2O2 in a NADPH-dependent manner, although with low catalytic efficiency. In the presence of thioredoxin, mitochondrial TGR showed a thioredoxin peroxidase-like activity. All disulfide reductase activities were inhibited by auranofin, suggesting mTGR is dependent on selenocysteine. The reductase activity with GSSG showed a higher dependence on temperature as compared with the DTNB reductase activity. The variation of the GSSG- and DTNB reductase activities on pH was dependent on the disulfide substrate. Like the cytosolic isoform, mTGR showed a hysteretic kinetic behavior at moderate or high GSSG concentrations, but it was less sensitive to calcium. The enzyme was able to protect glutamine synthetase from oxidative inactivation, suggesting that mTGR is competent to contend with oxidative stress.

Full Text Available Copper nanoflowers have been fabricated using two different techniques; electro-deposition of copper in polymer and anodic alumina templates, and cytyltrimethal ammonium bromide (CTAB-assisted hydrothermal method. Scanning Electron Microscope (SEM images record some interesting morphologies of metallic copper nanoflowers. Field Emission Scanning Electron Microscope (FESEM has been used to determine morphology and composition of copper oxide nanoflowers. X-ray diffraction (XRD pattern reveals the monoclinic phase of CuO in the crystallographic structure of copper oxide nanoflowers. There is an element of random artistic design of nature, rather than science, in exotic patterns of nanoflowers fabricated in our laboratory.

The results of the study demonstrated that the content of nitrates and nitrites in various baker's products varied from 0.96 (in wheat rolls and baguettes) to 44.07 mg KO3/kg in pumpernickel bread. In wholemeal bread, village bread, tourist bread, rye brown bread and Graham bread the content of these compounds was from 1.46 to 27.10 mg KNO3/kg. The mean content of nitrites in these bread sorts was 1.76 mg NaNo2/kg, range 0.10-4.40 mg NaNo2/kg. In white wheat flours (Wrocław flour, cake flour and Poznań flour) the content of nitrates ranged from 1.10 to 19.08 mg KNO3/kg, and in the dishes produced from them in household was from 0.50 to 16.33 mg KNO3/kg. The content of nitrites in these flours was in the range from 0.00 to 4.16 mg NaNo2/kg, and in the products prepared from them it was from 0.00 to 1.60 mg NaNO2/kg. Eleven types were tested also of popular biscuits, wafers, gingerbread and hard cakes in which the content of nitrates was from 3.66 to 17.72 mg/kg, and that of nitrites was from 0.00 to 8.80 mg NaNo2/kg. Considering the average consumption of these products per one person in the seashore region and the mean values of nitrates and nitrites it was calculated that they provided daily about 3.9 mg KNO3 and 0.4 NaNo2, that is about 1.8% of nitrates and 7.7% of nitrites consumed by adults in daily food ration.

Full Text Available The presented paper deals with the determination of nitrates and nitrites content in carrots, in different vegetation stages of the carrot culture. High nitrates and nitrites concentration in vegetables is mainly due to excessive nitrogen content in the soil system, thus deteriorating the nutritional and hygienic values of products and complicating the processing and storage. The determination was tested on carrot samples assayed from an experimental field set up near Timisoara. In experimental field, to the carrot culture was administrated different doses of fertilizers (NPK and the samples for analysis were assayed in different phases of vegetation. The obtained results indicated that the highest level of nitrate in carrots was found to the variant b3 (N150P90K90 in experimental field, who was above maximum limit allowed (LMA. Maximum limit allowed for nitrates in carrots, in accordance with ORDER No. 293/640/2001-1/2002 regarding security and quality conditions for vegetables and fresh fruits for human consumption is 400 ppm. For all other samples of carrots the nitrates level was below of LMA. The nitrite content grows in case of fertilizer administration during the whole vegetation stages of the plant. In variant N150P90K90 the nitrite content was above (LMA in carrot samples in all stages of vegetation. The nitrite content in carrots should not exceed 1-2 ppm. Nitrate and nitrite content in carrots was done with the help of High Performance Liquid Chromatography (HPLC in the Laboratory for the Measurement of Residues of the Department of Agro-techniques of the U.S.A-V.M.B in Timisoara.

Highlights: ► Nitrite enhanced the photo-damage by ZnO nanoparticles to BSA and HaCaT cells. ► Protein nitration was induced by nitrite in photo-damaged BSA and HaCaT cells. ► The effects of photo-damage on BSA were affected by various factors. ► 50-nm ZnO induced more apoptosis than 90-nm ZnO in HaCaT cells. -- Abstract: Zinc oxide nanoparticles are widely used in sunscreen products because of their chemical stability and capability of blocking harmful ultraviolet rays. However, zinc oxide nanoparticles can also generate reactive species under ultraviolet irradiation. Because nitrite can form reactive nitrogen species under oxidative stress and because it exists in perspiration and cosmetics, we studied the effects of nitrites on the photocatalytic damage of zinc oxide nanoparticles (50 nm and 90 nm) to bovine serum albumin and human keratinocyte cells under ultraviolet irradiation (365 nm and 254 nm). The results indicate that nitrite plays an enhancing role in photocatalytic damage by breaking amino acid residues and promoting protein oxidation and nitration. The concentrations of zinc oxide and nitrite, the irradiation light and duration, and the pH of the medium are important factors influencing this photocatalytic damage. Size effects of ZnO nanoparticles on bovine serum albumin and keratinocyte cells are different. It is speculated that the extent of photo-damage is partially dependent on the aggregation of zinc oxide. These findings may be valuable for understanding potential risks of applying zinc oxide nanoparticle-containing sunscreens to human skin under sunlight exposure.

Full Text Available Due to the negative impact of nitrate and nitrite on human health, their presence exceeding acceptable levels is not desired in foodstuffs. Thus, nitrite determination at low concentrations is a major challenge in electroanalytical chemistry, which can be achieved by fast, cheap, and safe electrochemical sensors. In this work, the working electrode (Au was functionalized with p-aminothiophenol (p-ATP and modified with gold nanoparticles (Au-NPs to manufacture the final (Au/p-ATP-Aunano electrode in a two-step procedure. In the first step, p-ATP was electropolymerized on the electrode surface to obtain a polyaminothiophenol (PATP coating. In the second step, Au/p-ATP-Aunano working electrode was prepared by coating the surface with the use of HAuCl4 solution and cyclic voltammetry. Determination of aqueous nitrite samples was performed with the proposed electrode (Au/p-ATP-Aunano using square wave voltammetry (SWV in pH 4 buffer medium. Characteristic peak potential of nitrite samples was 0.76 V, and linear calibration curves of current intensity versus concentration was linear in the range of 0.5–50 mg·L−1 nitrite with a limit of detection (LOD of 0.12 mg·L−1. Alternatively, nitrite in sausage samples could be colorimetrically determined with high sensitivity by means of p-ATP‒modified gold nanoparticles (AuNPs and naphthylethylene diamine as coupling agents for azo-dye formation due to enhanced charge-transfer interactions with the AuNPs surface. The slopes of the calibration lines in pure NO2− solution and in sausage sample solution, to which different concentrations of NO2− standards were added, were not significantly different from each other, confirming the robustness and interference tolerance of the method. The proposed voltammetric sensing method was validated against the colorimetric nanosensing method in sausage samples.

Due to the negative impact of nitrate and nitrite on human health, their presence exceeding acceptable levels is not desired in foodstuffs. Thus, nitrite determination at low concentrations is a major challenge in electroanalytical chemistry, which can be achieved by fast, cheap, and safe electrochemical sensors. In this work, the working electrode (Au) was functionalized with p-aminothiophenol (p-ATP) and modified with gold nanoparticles (Au-NPs) to manufacture the final (Au/p-ATP-Aunano) electrode in a two-step procedure. In the first step, p-ATP was electropolymerized on the electrode surface to obtain a polyaminothiophenol (PATP) coating. In the second step, Au/p-ATP-Aunano working electrode was prepared by coating the surface with the use of HAuCl₄ solution and cyclic voltammetry. Determination of aqueous nitrite samples was performed with the proposed electrode (Au/p-ATP-Aunano) using square wave voltammetry (SWV) in pH 4 buffer medium. Characteristic peak potential of nitrite samples was 0.76 V, and linear calibration curves of current intensity versus concentration was linear in the range of 0.5-50 mg·L(-1) nitrite with a limit of detection (LOD) of 0.12 mg·L(-1). Alternatively, nitrite in sausage samples could be colorimetrically determined with high sensitivity by means of p-ATP‒modified gold nanoparticles (AuNPs) and naphthylethylene diamine as coupling agents for azo-dye formation due to enhanced charge-transfer interactions with the AuNPs surface. The slopes of the calibration lines in pure NO₂(-) solution and in sausage sample solution, to which different concentrations of NO₂(-) standards were added, were not significantly different from each other, confirming the robustness and interference tolerance of the method. The proposed voltammetric sensing method was validated against the colorimetric nanosensing method in sausage samples.

Due to the negative impact of nitrate and nitrite on human health, their presence exceeding acceptable levels is not desired in foodstuffs. Thus, nitrite determination at low concentrations is a major challenge in electroanalytical chemistry, which can be achieved by fast, cheap, and safe electrochemical sensors. In this work, the working electrode (Au) was functionalized with p-aminothiophenol (p-ATP) and modified with gold nanoparticles (Au-NPs) to manufacture the final (Au/p-ATP-Aunano) electrode in a two-step procedure. In the first step, p-ATP was electropolymerized on the electrode surface to obtain a polyaminothiophenol (PATP) coating. In the second step, Au/p-ATP-Aunano working electrode was prepared by coating the surface with the use of HAuCl4 solution and cyclic voltammetry. Determination of aqueous nitrite samples was performed with the proposed electrode (Au/p-ATP-Aunano) using square wave voltammetry (SWV) in pH 4 buffer medium. Characteristic peak potential of nitrite samples was 0.76 V, and linear calibration curves of current intensity versus concentration was linear in the range of 0.5–50 mg·L−1 nitrite with a limit of detection (LOD) of 0.12 mg·L−1. Alternatively, nitrite in sausage samples could be colorimetrically determined with high sensitivity by means of p-ATP‒modified gold nanoparticles (AuNPs) and naphthylethylene diamine as coupling agents for azo-dye formation due to enhanced charge-transfer interactions with the AuNPs surface. The slopes of the calibration lines in pure NO2− solution and in sausage sample solution, to which different concentrations of NO2− standards were added, were not significantly different from each other, confirming the robustness and interference tolerance of the method. The proposed voltammetric sensing method was validated against the colorimetric nanosensing method in sausage samples. PMID:27490543

Full Text Available The zeolite with the unit formula of Na8(Al6Si6O24S.4H2O was synthesized directly on red mud base with addition of single silicon (signed as RH-ZeO-Si and both silicon and aluminum portions (signed as RM-ZeO-SiAl to original Tan Rai (Vietnam red mud. The structure of the zeolite was studied by X-Ray difration and FT-IR absorption spectra. The synthesized materials were studied on their adsorption ability of ammonium and nitrite ions. The results showed that, the adsorption of ammonium cation was mostly allowing ion-exchange mechanism and the zeolite crystaline forms played predominantly role besids minor one of single metal oxides. For nitrite anions, it is otherwise, the adsorption mechanism was mostly leant to electrostatic attraction between nitrite anions and electropositive effect of the hematite surface in light acidic condition. The adsorption isotherms of all ammonium and nitrite ions on both synthesized materials were nearly conformable with Freundlich model than Langmuir model. Those showed that, both materials have unhomogeneous adsorption surface. The maximum adsorption capacity of ammonium and nitrite on RM-ZeO-Si was 5.71 mg/g and 2.73 mg/g respectively, and on RM-ZeO-SiAl was 5.61 mg/g and 3.12 mg/g respectively. The initial test of competitive ions influencing on adsorption ability showed that, for all cases the competition of cations to ammonium ion was more significance than those of anions to nitrite ion in the same conditions.

Full Text Available Nitrates are the basic source of nitrogen for the majority of plants. Absorption and transformation of nitrates in plants are determined by external conditions and, first of all, temperature and light intensity. The influence of the temperature increasing till +40 0С on activity of nitrate reductase was studied. It is shown, that the rise of temperature was accompanied by sharp decrease of activity nitrate reductase in leaves of winter wheat, what, apparently, occurred for the account deactivations of enzyme and due to its dissociation.

The observation that oxygen represses nitrate reductase biosynthesis in a hemA mutant grown aerobically with or without delta-aminolevulinic acid indicates that cytochromes are not responsible for nitrate reductase repression in aerobically grown cells. PMID:326768

Full Text Available Copper is a transition metal that has been linked to pathological and beneficial effects in neurodegenerative diseases. In Parkinson’s disease, free copper is related to increased oxidative stress, alpha-synuclein oligomerization, and Lewy body formation. Decreased copper along with increased iron has been found in substantia nigra and caudate nucleus of Parkinson’s disease patients. Copper influences iron content in the brain through ferroxidase ceruloplasmin activity; therefore decreased protein-bound copper in brain may enhance iron accumulation and the associated oxidative stress. The function of other copper-binding proteins such as Cu/Zn-SOD and metallothioneins is also beneficial to prevent neurodegeneration. Copper may regulate neurotransmission since it is released after neuronal stimulus and the metal is able to modulate the function of NMDA and GABA A receptors. Some of the proteins involved in copper transport are the transporters CTR1, ATP7A, and ATP7B and the chaperone ATOX1. There is limited information about the role of those biomolecules in the pathophysiology of Parkinson’s disease; for instance, it is known that CTR1 is decreased in substantia nigra pars compacta in Parkinson’s disease and that a mutation in ATP7B could be associated with Parkinson’s disease. Regarding copper-related therapies, copper supplementation can represent a plausible alternative, while copper chelation may even aggravate the pathology.

Pitting corrosion could be extremely serious for dilute high-level radioactive waste stored or processed in carbon steel tanks at the Savannah River Site. In these solutions, nitrate is an aggressive ion with respect to pitting of carbon steel while nitrite can be used as an inhibitor. Excessive additions of nitrite increase the risk of generating unstable nitrogen compounds during waste processing, and insufficient additions of nitrite could increase the risk of corrosion-induced failure. Thus there are strong incentives to obtain a fundamental understanding of the role of nitrite in pitting corrosion prevention with these solution chemistries. In this dissertation, both a 1-D and a 2-D model are used to study the pitting mechanism as a function of nitrite/nitrate ratios. The 1-D model used BAND(J) to test a reaction mechanism for the passivation behavior by comparing the predicted Open Circuit Potential (OCP) with OCP data from experiments at different NO2-/NO3- ratio. The model predictions are compared with Cyclic Potentiodynamic Polarization (CPP) experiments. A 2-D model was developed for the propagation of a pit in iron by writing subroutines for finite element software of GAMBIT and FIDAP. Geometrically distributed anodic and cathodic reactions are assumed. The results show three partial explanations describing the inhibition influence of nitrite to iron corrosion: the competing reduction reaction of nitrate to nitrite, the formation of Fe(OH)+, and the function of the porous film. The current distributions and the effect of porosity of the film on pH are also explained. The calculation results also show that rate of pit growth decreases as the pit diameter increases until it reaches a constant value. The profile of the local current density on the pit wall is parabolic for small pits and it changes to a linear distribution for large pits. The model predicts that addition of nitrite will decrease the production of ferrous ions and those can prevent iron from

Corrosion of rebar steel due to environmental causes has been studied through various approaches, and among the protection techniques use of inhibitors has gained encouragement. Nitrites and nitrates of sodium have gained sufficient scientific coverage. Recently, nitrites and nitrates of calcium have been verified in some studies, which, however, needs further experimentation through different angles. Simple polarization technique has been utilized in the present study to compare inhibitive efficiency of these salts of sodium and calcium, which indicate that calcium salts are more efficient.

The nitrate and nitrite contents in cheese of the Gouda type, produced by the Milk Complex of Havanna, were investigated and compared with those found in 5 types of imported cheese consumed in Cuba. No significant differences were found in the mean value (mean) of NaNO3, whereas in the mean value (mean) of NaNO2 differences were found when comparing all cheeses against each other. From these results it is inferred that the consumption of Gouda type cheese does not represent an important contribution to the daily intake of nitrates and nitrites, precursors in the synthesis of cancerogenic N-nitroso compounds.

Uranium terminal mono-oxo complexes are prepared with a unique activation of nitrite following reductive cleavage of an N-O bond with loss of nitric oxide. The thermodynamic driving force of U═O bond formation differentiates this reactivity from known mechanisms of nitrite reduction, which are typically mediated by proton transfer. Mechanistic details are explored by DFT supporting a simple homolytic cleavage pathway from a κ(1)-ONO bound intermediate. Complexes of the formula U(VI)OX[N(SiMe(3))(2)](3) are formed providing a trigonal bipyramidal framework into which ligands trans to the U═O bond may be installed.

Iron-doped titania nanoparticles exhibit a higher photocatalytic activity than pure TiO2 for the degradation of nitrite. The optimum Fe-doped content in terms of activity is approximately 0. 5 %. The increase in photoactivity is probably due to the higher adsorption and the inhibition of electron-hole recombination. The photocatalytic oxidation reaction of nitrite over the Fe-doped TiO2 catalyst follows zero-order kinetics, which is different from that over pure TiO2. The reaction rate decreases linearly with the increase of the pH of the solution.

Copper electrolyte was purified by copper arsenite that was prepared with As2O3. And electrolysis experiments of purified electrolyte were carried out at 235 and 305 A/m2, respectively. The results show that the yield of copper arsenite is up to 98.64% when the molar ratio of Cu to As is 1.5 in the preparation of copper arsenite. The removal rates of Sb and Bi reach 74.11% and 65.60% respectively after copper arsenite is added in electrolyte. The concentrations of As, Sb and Bi in electrolyte nearly remain constant during electrolysis of 13 d. The appearances of cathode copper obtained at 235 and 305 A/m2 are slippery and even, and the qualification rate is 100% according to the Chinese standard of high-pure cathode copper(GB/T467-97).

For 422 male patients with symptoms indicative of a urinary tract infection, nitrite and leukocyte esterase activity dipstick test results were compared with results of culture of urine samples. The positive predictive value of a positive nitrite test result was 96%. Addition of results of the leuko

Impact of nitrite on aerobic phosphorus (P) uptake of poly-phosphate accumulating organisms (PAOs) in three different enhanced biological phosphorus removal (EBPR) systems was investigated, i.e., the enriched PAOs culture fed with synthetic wastewater, the two lab-scale sequencing batch reactors (SBRs) treating domestic wastewater for nutrient removal through nitrite-pathway nitritation and nitrate-pathway nitrification, respectively. Fluorescence in situ hybridization results showed that PAOs in the three sludges accounted for 72, 7.6 and 6.5% of bacteria, respectively. In the enriched PAOs culture, at free nitrous acid (FNA) concentration of 0.47 × 10(-3) mg HNO₂-N/L, aerobic P-uptake and oxidation of intercellular poly-β-hydroxyalkanoates were both inhibited. Denitrifying phosphorus removal under the aerobic conditions was observed, indicating the existence of PAOs using nitrite as electron acceptor in this culture. When the FNA concentration reached 2.25 × 10(-3) mg HNO2-N/L, denitrifying phosphorus removal was also inhibited. And the inhibition ceased once nitrite was exhausted. Corresponding to both SBRs treating domestic wastewater with nitritation and nitrification pathway, nitrite inhibition on aerobic P-uptake by PAOs did not occur even though FNA concentration reached 3 × 10(-3) and 2.13 × 10(-3) mg HNO₂-N/L, respectively. Therefore, PAOs taken from different EBPR activated sludges had different tolerance to nitrite.

For 422 male patients with symptoms indicative of a urinary tract infection, nitrite and leukocyte esterase activity dipstick test results were compared with results of culture of urine samples. The positive predictive value of a positive nitrite test result was 96%. Addition of results of the

Breaking the rules: Reversal of the high Markovnikov selectivity of Wacker-type oxidations was accomplished using a nitrite co-catalyst. Unbiased aliphatic alkenes can be oxidized with high yield and aldehyde selectivity, and several functional groups are tolerated. 18O-labeling experiments indicate that the aldehydic O atom is derived from the nitrite salt.

Nitrite (NO2-) functions as an important nitric oxide (NO) donor under hypoxic conditions.Both nitrite and NO have been found to protect the mammalian heart and other tissues against ischemia(anoxia)-reoxygenation injury by interacting with mitochondrial electron transport complexes and limiting ...

We report on the photodissociation dynamics study of n-butyl nitrite (n-C_4H_9ONO) and isoamyl nitrite ((CH_3)_2C_3H_5ONO) by means of time-resolved Fourier transform infrared (TR-FTIR) emission spectroscopy. The obtained TR-FTIR emission spectra of the nascent NO fragments produced in the 355-nm laser photolysis of the two alkyl nitrite species showed an almost identical rotational temperature and vibrational distributions of NO. In addition, a close resemblance between the two species was also found in the measured temporal profiles of the IR emission of NO and the recorded UV absorption spectra. The experimental results are consistent with our ab initio calculations using the time-dependent density functional theory at the B3LYP/6-311G(d,p) level, which indicate that the substitution of one of the two {gamma}-H atoms in n-C_4H_9ONO with a methyl group to form (CH_3)_2C_3H_5ONO has only a minor effect on the photodissociation dynamics of the two molecules.

The effect of the simultaneous exposure to transition metals and natural antioxidants frequently present in food is a question that needs further investigation. We aimed to explore the possible use of the natural polyphenols caffeic acid (CA), resveratrol (RES) and curcumin (CUR) to prevent damages induced by copper-overload on cellular molecules in HepG2 and A-549 human cells in culture. Exposure to 100μM/24h copper (Cu) caused extensive pro-oxidative damage evidenced by increased TBARS, protein carbonyls and nitrite productions in both cell types. Damage was aggravated by simultaneous incubation with 100μM of CA or RES, and it was also reflected in a decrease on cellular viability explored by trypan blue dye exclusion test and LDH leakage. Co-incubation with CUR produced opposite effects demonstrating a protective action which restored the level of biomarkers and cellular viability almost to control values. Thus, while CA and RES might aggravate the oxidative/nitrative damage of Cu, CUR should be considered as a putative protective agent. These results could stimulate further research on the possible use of natural polyphenols as neutralizing substances against the transition metal over-exposure in specific populations such as professional agrochemical sprayers and women using Cu-intrauterine devices.

higher (p<0.05) in the broodfish fed CSD0 and CSD1 diets than the other diets. Exposure of Clarias gariepinus fish to copper in water, at concentrations above 1.0mg CuSO4/g elicits adverse ... introduction of a toxicant to an aquatic system ..... Toxicity of four commonly used agrochemicals on. Oreochromis niloticus (L) fry.

The adulteration of urine specimens with nitrite ion hasseen shown to mask the gas chromatography-mass spectrometry (GC-MS) confirmation testing of marijuana use. This study was designed to further investigate the effect of nitrite adulteration on the detection of five commonly abused drugs by immunoassay screening and GC-MS analysis. The drugs tested are cocaine metabolite (benzoylecgonine), morphine, 11-nor-delta-tetrahydrocannabinol-9-carboxylic acid (THCCOOH), amphetamine, and phencyclidine. The immunoassays evaluated included the instrument-based Abuscreen ONLINE assays, the on-site Abuscreen ONTRAK assays, and the one-step ONTRAK TESTCUP-5 assay. Multianalyte standards containing various levels of drugs were used to test the influence of both potassium and sodium nitrite. In the ONLINE immunoassays, the presence of up to 1.0M nitrite in the multianalyte standards had no significant effect for benzoylecgonine, morphine, and phencyclidine assays. With a high concentration of nitrite, ONLINE became more sensitive for amphetamine (detected more drug than what was expected) and less sensitive for THCCOOH (detected less drug than what was expected). No effects of nitrite were observed on the results of the Abuscreen ONTRAK assays. Similarly, no effects were observed on the absolute qualitative results of the TESTCUP-5 when testing the nitrite-adulterated standards. However, the produced intensities of the signals that indicate the negative test results were slightly lowered in the THC and phencyclidine assays. The presence of 1.0M of nitrite did not show dramatic interference with the GC-MS analysis of benzoylecgonine, morphine, amphetamine, and phencyclidine. In contrast, nitrite ion significantly interfered with the detection of THCCOOH by GC-MS. The presence of 0.03M of nitrite ion resulted in significant loss in the recovery of THCCOOH and its internal standard by GC-MS. The problem of nitrite adulteration could be alleviated by sodium bisulfite treatment even

The activity of fumaric reductase in Cysticercus cellulosae tissue homogenate with albendazole and oxfendazole individually was detected. Results showed that the two kinds of drugs both could inhabite the activity of fumaric reductase. The results indicate that the mechanism of action of benzimidazole carbamate drugs is probably inhabiting the complex of fumaric reductase noncompetently, thus lead to the exhaostion of energy and death.

Full Text Available Dietary copper is essential for multicellular organisms. Copper is redox active and required as a cofactor for enzymes such as the antioxidant Superoxide Dismutase 1 (SOD1. Copper dyshomeostasis has been implicated in Alzheimer's disease. Mutations in the presenilin genes encoding PS1 and PS2 are major causes of early-onset familial Alzheimer's disease. PS1 and PS2 are required for efficient copper uptake in mammalian systems. Here we demonstrate a conserved role for presenilin in dietary copper uptake in the fly Drosophila melanogaster. Ubiquitous RNA interference-mediated knockdown of the single Drosophila presenilin (PSN gene is lethal. However, PSN knockdown in the midgut produces viable flies. These flies have reduced copper levels and are more tolerant to excess dietary copper. Expression of a copper-responsive EYFP construct was also lower in the midgut of these larvae, indicative of reduced dietary copper uptake. SOD activity was reduced by midgut PSN knockdown, and these flies were sensitive to the superoxide-inducing chemical paraquat. These data support presenilin being needed for dietary copper uptake in the gut and so impacting on SOD activity and tolerance to oxidative stress. These results are consistent with previous studies of mammalian presenilins, supporting a conserved role for these proteins in mediating copper uptake.

-through biofilm system to continuous immigration from a tap water metacommunity while applying different nitrite surface loading rates. After 63 days of operation, we extracted biofilms and analyzed the community composition via Illumina MiSeq targeting the 16S rRNA gene. Previous studies have shown......Four main processes are considered to drive microbial community assembly: selection, drift, dispersal and speciation. These processes occur simultaneously, but the extent to which each process contributes to community assembly is unclear in natural communities. We exposed a high-throughput flow...... that Nitrospira is the dominant nitrite oxidizing genus in low nitrite environments. Hence, we postulated that by elevating the nitrite surface loading we would select for NOB with lower nitrite affinity than Nitrospira. We observed different dominant NOB species under different loading rates. While...

A structural and functional model of bacterial nitric oxide reductase (NOR) has been designed by introducing two glutamates (Glu) and three histidines (His) in sperm whale myoglobin. X-ray structural data indicate that the three His and one Glu (V68E) residues bind iron, mimicking the putative FeB site in NOR, while the second Glu (I107E) interacts with a water molecule and forms a hydrogen bonding network in the designed protein. Unlike the first Glu (V68E), which lowered the heme reduction potential by {approx}110 mV, the second Glu has little effect on the heme potential, suggesting that the negatively charged Glu has a different role in redox tuning. More importantly, introducing the second Glu resulted in a {approx}100% increase in NOR activity, suggesting the importance of a hydrogen bonding network in facilitating proton delivery during NOR reactivity. In addition, EPR and X-ray structural studies indicate that the designed protein binds iron, copper, or zinc in the FeB site, each with different effects on the structures and NOR activities, suggesting that both redox activity and an intermediate five-coordinate heme-NO species are important for high NOR activity. The designed protein offers an excellent model for NOR and demonstrates the power of using designed proteins as a simpler and more well-defined system to address important chemical and biological issues.

The enzyme nitrogenase complex is a key component conferring nitrogen fixation in all known diazotrophs. This study for the first time examines the impact of As, Na, Cd, Cu and butachlor on component II (dinitrogenase reductase, nifH1) of nitrogenase from diazotrophic cyanobacterium Anabaena sp. PCC7120 using in silico and wet lab approaches. The nifH1 of Anabaena is a glycine-rich stable protein having DNA-binding properties and shows close similarity with free living compared with symbiotic diazotrophs. Phylogenetic tree revealed an adverse effect of the selected stresses on close homologs across the diazotroph community. The protein interaction network demonstrated the presence of nirA, glnA, glnB, alr4255 and alr2485 proteins besides nif proteins, suggesting their involvement in nitrogen fixation along with nifH1. Homology modelling and docking under As, Na, Cd, Cu and butachlor revealed an interaction between stressors and nifH1 protein which was further validated by a transcript of the gene through quantitative real-time PCR (qRT-PCR). Presence of binding sites for As, Na, Cd and Cu on oxyR promoter attested their adverse affects on nifH1. Maximum down-regulation of nifH1 in Cd and As followed by salt, copper and butachlor revealed that arsenic and cadmium were most potential inhibitors of nitrogenase of diazotrophic community, which might negatively affect crop yield.

Understanding the origin and evolution of cellular processes is fundamental to understand how biological activity has shaped the history of our planet. Among these, aerobic respiration is probably one of the most debated. We have applied a phylogenomics approach to investigate the origin and evolution of dioxygen reductases (O(2)Red), the key enzymes of aerobic respiratory chains. The distribution and phylogenetic analysis of the four types of O(2)Red (Cyt-bd and the A, B, and C families of heme-copper O(2)Red) from 673 complete bacterial and archaeal genomes show that these enzymes have very different evolutionary histories: Cyt-bd are of bacterial origin and were transferred to a few archaea; C-O(2)Red are of proteobacterial origin and were transferred to a few other bacteria; B-O(2)Red are of archaeal origin and were transferred to a few bacteria; and A-O(2)Red are the most ancient O(2)Red and were already present prior to the divergence of major present-day bacterial and archaeal phyla, thus before the emergence of Cyanobacteria and oxygenic photosynthesis. Implications for the origin and the evolution of aerobic respiration are discussed.

A structural and functional model of bacterial nitric oxide reductase (NOR) has been designed by introducing two glutamates (Glu) and three histidines (His) in sperm whale myoglobin. X-ray structural data indicate that the three His and one Glu (V68E) residues bind iron, mimicking the putative FeB site in NOR, while the second Glu (I107E) interacts with a water molecule and forms a hydrogen bonding network in the designed protein. Unlike the first Glu (V68E), which lowered the heme reduction potential by {approx}110 mV, the second Glu has little effect on the heme potential, suggesting that the negatively charged Glu has a different role in redox tuning. More importantly, introducing the second Glu resulted in a {approx}100% increase in NOR activity, suggesting the importance of a hydrogen bonding network in facilitating proton delivery during NOR reactivity. In addition, EPR and X-ray structural studies indicate that the designed protein binds iron, copper, or zinc in the FeB site, each with different effects on the structures and NOR activities, suggesting that both redox activity and an intermediate five-coordinate heme-NO species are important for high NOR activity. The designed protein offers an excellent model for NOR and demonstrates the power of using designed proteins as a simpler and more well-defined system to address important chemical and biological issues.

Copper, though essential, is potentially toxic heavy metal at supraoptimal level and has widespread contamination. The present investigation was carried out to study the responses induced by lower as well as higher doses of copper (0.1-25 microM) in an aquatic macrophyte, Hydrilla verticillata (L.f.) Royle for a period of 1-7 days. The plants accumulated copper in high amount with a maximum of 770 microg g(-1) dw on day 7 at 25 microM. Biomass and photosynthetic pigments showed less alteration up to 1 microM while at higher concentrations, significant decline occurred. Malondialdehyde (MDA) content and electrical conductivity (EC) also showed sharp increase at higher concentrations indicating oxidative stress. In response to copper exposure, plants showed significant induction of proteins and enzymes like superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), catalase (CAT) and glutathione reductase (GR), however, only up to moderate exposures. Total non-protein thiols (NP-SH) and cysteine levels increased significantly up to 5 microM copper exposure while at 25 microM, their level declined drastically. Reduced glutathione (GSH) showed decrease at all concentrations while oxidized glutathione (GSSG) simultaneously increased. Phytochelatins (PCs) were also induced significantly at studied concentrations of 1 and 5 microM on day 4 in comparison to control. However, copper chelation depicted by PC-SH to copper ratio was found to be low (6.5% at 1 microM and 2.4% at 5 microM) suggesting that PCs play only a part in integrated mechanisms of copper homeostasis and detoxification. Tolerant response of plants to moderate copper exposures and high accumulation potential warrants their suitability for remediation of moderately copper polluted water bodies.

Full Text Available Ceruloplasmin, the main copper binding protein in blood plasma, has been of particular interest for its role in efflux of iron from cells, but has additional functions. Here we tested the hypothesis that it releases its copper for cell uptake by interacting with a cell surface reductase and transporters, producing apoceruloplasmin. Uptake and transepithelial transport of copper from ceruloplasmin was demonstrated with mammary epithelial cell monolayers (PMC42 with tight junctions grown in bicameral chambers, and purified human (64Cu-labeled ceruloplasmin secreted by HepG2 cells. Monolayers took up virtually all the (64Cu over 16h and secreted half into the apical (milk fluid. This was partly inhibited by Ag(I. The (64Cu in ceruloplasmin purified from plasma of (64Cu-injected mice accumulated linearly in mouse embryonic fibroblasts (MEFs over 3-6h. Rates were somewhat higher in Ctr1+/+ versus Ctr1-/- cells, and 3-fold lower at 2 °C. The ceruloplasmin-derived (64Cu could not be removed by extensive washing or trypsin treatment, and most was recovered in the cytosol. Actual cell copper (determined by furnace atomic absorption increased markedly upon 24h exposure to holoceruloplasmin. This was accompanied by a conversion of holo to apoceruloplasmin in the culture medium and did not occur during incubation in the absence of cells. Four different endocytosis inhibitors failed to prevent 64Cu uptake from ceruloplasmin. High concentrations of non-radioactive Cu(II- or Fe(III-NTA (substrates for cell surface reductases, or Cu(I-NTA (to compete for transporter uptake almost eliminated uptake of (64Cu from ceruloplasmin. MEFs had cell surface reductase activity and expressed Steap 2 (but not Steaps 3 and 4 or dCytB. However, six-day siRNA treatment was insufficient to reduce activity or uptake. We conclude that ceruloplasmin is a circulating copper transport protein that may interact with Steap2 on the cell surface, forming apoceruloplasmin, and Cu(I that

Disorders of copper metabolism are associated with neurological dysfunction including Wilson's disease (WD). WD is a autosomal recessive disorder caused by mutations in the ATP7B gene resulting in the inability of the hepatocytes to remove excess copper. Gradual copper accumulation causes damage to liver, brain and other organs manifesting in liver disease, neurological and psychiatric symptoms. Also scond copper-neurometaboic disorder: Menkes disease charaterized with mutated ATP7A gene, is ralated with abnormally neuroal transmission and synaptogenesis. Parkinson's disease and Alzheimer's disease both are refered to some degree of copper/iron metabolism changes. The precise mechanisms by which excess copper causes neurological damage remain to be elucidated. In this study, we aimed to investigate the influence of excessive amounts of Cu(2+) on the oxidative damage response and survival of primary astrocytes from newborn rats. Primary cultured rat astrocytes were divided into three groups: 30 μmol/L CuCl2, 100 μmol/L CuCl2 and control. At 12, 24, 48, 96 and 120 hours of CuCl2 intervention, cell viability, intracellular reduced glutathione level and glutathion reductase activity, and nitric oxide secretion were determined. It was found that 30 μmol/L CuCl2 might stimulate the exaltation and the compensatory proliferation of astrocytes. The survival rate of astrocytes in the 100 μmol/L CuCl2 group was significantly decreased relative to the 30 μmol/L CuCl2 group. At 24 hours of CuCl2 intervention, intracellular reduced glutathione level and glutathion reductase activity were significantly decreased in the 100 μmol/L CuCl2 group compared to the control group. At 120 hours of CuCl2 intervention, nitric oxide secretion in the 100 μmol/L CuCl2 group was significantly greater than in the control group. Under pathological conditions, excessive amounts of Cu(2+) greatly damaged the growth and proliferation of astrocytes, reduced the anti-oxidative capacity of

Denitrifying phosphorus removal (DPR) by denitrifying phosphorus-accumulating organisms (DPAOs) is a promising approach for reducing energy and carbon usage. However, influent fluctuations or interruptions frequently expose the DPAOs biomass to starvation conditions, reducing biomass activity and amount, and ultimately degrading the performance of DPR. Therefore, a better understanding of the endogenous metabolism and recovery ability of DPAOs is urgently required. In the present study, anaerobic starvation (12 days) and recovery were investigated in nitrite- and nitrate-cultivated DPAOs at 20 ± 1 °C. The cell decay rates in nitrite-DPAOs sludges from the end of the anaerobic and aerobic phase were 0.008 day⁻¹ and 0.007 day⁻¹, respectively, being 64% and 68% lower than those of nitrate-DPAOs sludges. Nitrite-DPAOs sludges also recovered more rapidly than nitrate-DPAOs sludge after 12 days of starvation. The maintenance energy of nitrite-DPAOs sludges from the end of the anaerobic and aerobic phase were approximately 31% and 34% lower, respectively, than those of nitrate-DPAOs sludges. Glycogen and polyphosphate (poly-P) sequentially served as the main maintenance energy sources in both nitrite-and nitrate-DPAOs sludges. However, the transformation pathway of the intracellular polymers during starvation differed between them. Nitrate-DPAOs sludge used extracellular polymeric substances (EPS) (mainly polysaccharides) as an additional maintenance energy source during the first 3 days of starvation. During this phase, EPS appeared to contribute to 19-27% of the ATP production in nitrate-DPAOs, but considerably less to the cell maintenance of nitrite-DPAOs. The high resistance of nitrite-DPAOs to starvation might be attributable to frequent short-term starvation and exposure to toxic substances such as nitrite/free nitrous acids in the parent nitrite-fed reactor. The strong resistance of nitrite-DPAOs sludge to anaerobic starvation may be exploited in P removal

An up-flow sludge blanket(UASB) and sequencing batch reactor(SBR) system was introduced to remove organics and nitrogen from landfill leachate.The synergetic effect of free ammonia(FA) inhibition and process control was used to achieve the nitrite pathway in the SBR.In previous research,inhibition of FA on nitrite oxidizing bacteria(NOB) activity has been revealed and the process control parameters(DO,ORP and pH) exactly indicate the end-point of nitritation.The method was implemented in the SBR achieving stable nitrogen removal via the nitrite pathway from landfill leachate.The degree of nitrite accumulation during the nitritation was monitored along with the simultaneous and advanced removal of organics and nitrogen in the UASB-SBR system.The nitrifying bacteria community was quantitatively analyzed by fluorescence in situ hybridization(FISH) techniques.Batch tests were carried out to investigate the denitritation kinetics of microbial bacteria in the SBR.Experimental results showed that the nitrite pathway could be repeatedly and reliably achieved by synergetic effect of FA inhibition and process control.FISH analysis showed the dominant nitrifying bacteria were ammonia-oxidizing β-Proteobacteria. Relationship between nitrite concentration and nitrite reduction rate followed the Monod-type equation.The maximum specific nitrite utilization rate(k) and half-velocity constant(Ks) were calculated as 0.44 gN gVSS-1d-1and 15.8 mg L-1,respectively.

Pneumocystis carinii dihydrofolate reductase (DHFR; 5,6,7,8-tetrahydrofolate: NADP+ oxidoreductase, EC 1.5.1.3) cDNA sequences have been isolated by their ability to confer trimethoprim resistance to Escherichia coli. Consistent with the recent conclusion that P. carinii is a member of the Fungi...

) reductases reported previously. Downstream of the butA gene of L. pseudomesenteroides, but coding in the opposite orientation, a putative DNA recombinase was identified. A two-step PCR approach was used to construct FPR02, a butA mutant of the wild-type strain, CHCC2114. FPR02 had significantly reduced...

Objective: Sepiapterin reductase deficiency (SRD) is an under-recognized levodopa-responsive disorder. We describe clinical, biochemical, and molecular findings in a cohort of patients with this treatable condition. We aim to improve awareness of the phenotype and available diagnostic and therapeuti

Dark-grown carrot (Daucus carota L.) tissue cultures were found to contain both protein components of the NADP/thioredoxin system--NADP-thioredoxin reductase and the thioredoxin characteristic of heterotrophic systems, thioredoxin h. Thioredoxin h was purified to apparent homogeneity and, like typical bacterial counterparts, was a 12-kdalton (kDa) acidic protein capable of activating chloroplast NADP-malate dehydrogenase (EC 1.1.1.82) more effectively than fructose-1,6-bisphosphatase (EC 3.1.3.11). NADP-thioredoxin reductase (EC 1.6.4.5) was partially purified and found to be an arsenite-sensitive enzyme composed of two 34-kDa subunits. Carrot NADP-thioredoxin reductase resembled more closely its counterpart from bacteria rather than animal cells in acceptor (thioredoxin) specificity. Upon greening of the cells, the content of NADP-thioredoxin-reductase activity, and, to a lesser extent, thioredoxin h decreased. The results confirm the presence of a heterotrophic-type thioredoxin system in plant cells and raise the question of its physiological function.

The paper reports the results of a comparative study of copper and copper halide vapor lasers emitting in a repetitively-pulsed regime. Copper chloride and copper bromide vapor lasers are found to have identical lasing characteristics under any excitation conditions. These characteristics are different from those of a copper vapor laser. An average lasing power of 13 W has been obtained for all lasers studied for an efficiency of 1%. It is shown that the choice of a laser will largely depend on the laser design suitability for a specific application.

The ascorbate-glutathione cycle is a metabolic pathway that detoxifies hydrogen peroxide and involves enzymatic and non-enzymatic antioxidants. Proteomic studies have shown that some enzymes in this cycle such as ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), and glutathione reductase (GR) are potential targets for post-translational modifications (PMTs) mediated by nitric oxide-derived molecules. Using purified recombinant pea peroxisomal MDAR and cytosolic and chloroplastic GR enzymes produced in Escherichia coli, the effects of peroxynitrite (ONOO(-)) and S-nitrosoglutathione (GSNO) which are known to mediate protein nitration and S-nitrosylation processes, respectively, were analysed. Although ONOO(-) and GSNO inhibit peroxisomal MDAR activity, chloroplastic and cytosolic GR were not affected by these molecules. Mass spectrometric analysis of the nitrated MDAR revealed that Tyr213, Try292, and Tyr345 were exclusively nitrated to 3-nitrotyrosine by ONOO(-). The location of these residues in the structure of pea peroxisomal MDAR reveals that Tyr345 is found at 3.3 Å of His313 which is involved in the NADP-binding site. Site-directed mutagenesis confirmed Tyr345 as the primary site of nitration responsible for the inhibition of MDAR activity by ONOO(-). These results provide new insights into the molecular regulation of MDAR which is deactivated by nitration and S-nitrosylation. However, GR was not affected by ONOO(-) or GSNO, suggesting the existence of a mechanism to conserve redox status by maintaining the level of reduced GSH. Under a nitro-oxidative stress induced by salinity (150mM NaCl), MDAR expression (mRNA, protein, and enzyme activity levels) was increased, probably to compensate the inhibitory effects of S-nitrosylation and nitration on the enzyme. The present data show the modulation of the antioxidative response of key enzymes in the ascorbate-glutathione cycle by nitric oxide (NO)-PTMs, thus indicating the close involvement of

We present a psychotropic bacteria–based biosensor that can be used in low–temperature seawater for the analysis of nitrate + nitrite (NOx –). The sensor can be used to resolve concentrations below 1 µmol L–1 at low temperature (<2.5°C) and high salinity (35‰), and in situ use in the deep sea was...

Currently, nitritation-anammox (anaerobic ammonium oxidation) bioreactors are designed to treat wastewaters with high ammonium concentrations at mesophilic temperatures (25 to 40°C). The implementation of this technology at ambient temperatures for nitrogen removal from municipal wastewater followin

The role played by curing agents (nitrite, ascorbate) on protein oxidation and Strecker aldehyde formation is studied. To fulfill this objective, increasing concentrations of nitrite (0, 75 and 150ppm) and ascorbate (0, 250 and 500ppm) were added to sausages subjected to a 54day drying process. The concurrence of intense proteolysis, protein carbonylation and formation of Strecker aldehydes during processing of sausages suggests that α-aminoadipic semialdehyde (AAS) and γ-glutamic semialdehyde (GGS) may be implicated in the formation of Strecker aldehydes. The fact that nitrite (150ppm, ingoing amount) significantly promoted the formation of protein carbonyls at early stages of processing and the subsequent formation of Strecker aldehydes provides strength to this hypothesis. Ascorbate (125 and 250ppm) controlled the overall extent of protein carbonylation in sausages without declining the formation of Strecker aldehydes. These results may contribute to understanding the chemistry fundamentals of the positive influence of nitrite on the flavor and overall acceptability of cured muscle foods.

Nitrate and nitrite contamination of surface waters (e.g. lakes) has become a severe environmental and health problem, especially in developing countries. The recent demonstration of nitrate reduction at the cathode of microbial fuel cell (MFC) provides an opportunity to develop a new technology ...

..., Deputy Agency Ethics Official, at 202-205-3088. Limited disclosure of business proprietary information... countervailing duties). If you are a trade/business association, provide the information, on an aggregate basis... determine whether revocation of the antidumping and countervailing duty orders on sodium nitrite from China...

Literatures revealed that the electron acceptor-nitrite could be inhibitory or toxic in the denitrifying phosphorus removal process.Batch test experiments were used to investigate the inhibitory effect during the anoxic condition.The inoculated activated sludge was taken from a continuous double- sludge denitrifying phosphorus and nitrogen removal system.Nitrite was added at the anoxic stage.One time injection and sequencing batch injection were carried on in the denitrifying dephosphorus procedure.The results indicated that the nitrite concentration higher than 30 mg/L would inhibit the anoxic phosphate uptake severely, and the threshold inhibitory concentration was dependent on the characteristics of the activated sludge and the operating conditions; instead, lower than the inhibitory concentration would not be detrimental to anoxic phosphorus uptake, and it could act as good electron acceptor for the anoxic phosphate accumulated.Positive effects performed during the denitrifying biological dephosphorus all the time.The utility of nitrite as good electron acceptor would provide a new feasible way in the denitrifying phosphorus process.

Some reactions (e.g., oxidation of nitrite, denitrification of ammonium) are accelerated in freeze-concentrated solution (FCS) compared to those in aqueous solution. Ice is highly intolerant to impurities, and the ice excludes those that would accelerate reactions. Here we show the acceleration of the N-nitrosation reaction of dimethylamine (DMA) with nitrite to produce N-nitrosodimethylamine (NDMA) in FCS. NDMA is a carcinogenic compound, and this reaction is potentially accelerated in frozen fish/meat. The eaction rate of the N-nitrosation reaction becomes fastest at specific pH. This means that it is a third-order reaction. Theoretical pH values of the peak in the third-order reaction are higher than the experimental one. Freeze-concentration of acidic solution causes pH decrement; however, the freeze-concentration alone could not explain the difference of pH values. The theoretical value was obtained under the assumption that no solute took part in ice. However, solutes are incorporated in ice with a small distribution coefficient of solutes into ice. This small incorporation enhanced the decrement of pH values. Using the distribution coefficient of chloride and sodium ion and assuming those of nitrite and DMA to explain the enhancement, we succeeded in estimating the distribution coefficients of nitrite: 2 × 10(-3) and DMA: 3 × 10(-2).

We present a new chemistry to determine nitrites implemented in a microfluidic paper-based analytical device (µPAD). The device is fabricated in cellulose paper with a sample reception area and three replicate detection areas with recognition chemistry immobilized by adsorption. The method involves the use of nitrite in an acid medium reaction to generate nitrous acid, which produces the oxidation of s-dihydrotetrazine: 1,2-dihydro-3,6-bis(3,5-dimethyl-1H-pyrazol-1-yl)-1,2,4,5-tetrazine (DHBPTz), which change the detection zone from colorless to pink. We used a digital camera and smartphone for the quantitative analysis of nitrite with the color coordinate S of the HSV color space as the analytical parameter. Parameters such as concentration and volume of s-dihydrotetrazine, pH, sample volume and reaction time were studied. The detection limit for this method is 1.30µM nitrite. To estimate the selectivity of the method an interference study of common ions in water samples was performed. The procedure was applied to natural water and compared with reference procedures.

Full Text Available This work reported new voltammetric/amperometric-based protocols using a commercial boron-doped diamond (BDD electrode for simple and fast simultaneous detection of sulfide and nitrite from water. Square-wave voltammetry operated under the optimized working conditions of 0.01 V step potential, 0.5 V modulation amplitude and 10 Hz frequency allowed achieving the best electroanalytical parameters for the simultaneous detection of nitrite and sulfide. For practical in-field detection applications, the multiple-pulsed amperometry technique was operated under optimized conditions, i.e., −0.5 V/SCE for a duration of 0.3 s as conditioning step, +0.85 V/SCE for a duration of 3 s that assure the sulfide oxidation and +1.25 V/SCE for a duration of 0.3 s, where the nitrite oxidation occurred, which allowed the simultaneously detection of sulfide and nitrite without interference between them. Good accuracy was found for this protocol in comparison with standardized methods for each anion. Also, no interference effect was found for the cation and anion species, which are common in the water matrix.

This work reported new voltammetric/amperometric-based protocols using a commercial boron-doped diamond (BDD) electrode for simple and fast simultaneous detection of sulfide and nitrite from water. Square-wave voltammetry operated under the optimized working conditions of 0.01 V step potential, 0.5 V modulation amplitude and 10 Hz frequency allowed achieving the best electroanalytical parameters for the simultaneous detection of nitrite and sulfide. For practical in-field detection applications, the multiple-pulsed amperometry technique was operated under optimized conditions, i.e., −0.5 V/SCE for a duration of 0.3 s as conditioning step, +0.85 V/SCE for a duration of 3 s that assure the sulfide oxidation and +1.25 V/SCE for a duration of 0.3 s, where the nitrite oxidation occurred, which allowed the simultaneously detection of sulfide and nitrite without interference between them. Good accuracy was found for this protocol in comparison with standardized methods for each anion. Also, no interference effect was found for the cation and anion species, which are common in the water matrix. PMID:26102487

Submerged attached growth bioreactors (SAGBs) were operated at 20 °C for 30 weeks in smart-aerated, partial nitritation ANAMMOX mode and in a timer-controlled, cyclic aeration mode. The smart-aerated SAGBs removed 48-53% of total nitrogen (TN) compared to 45% for SAGBs with timed aeration. Low dissolved oxygen concentrations and cyclic pH patterns in the smart-aerated SAGBs suggested conditions favorable to partial nitritation ANAMMOX and stoichiometrically-derived and numerically modeled estimations attributed 63-68% and 14-44% of TN removal to partial nitritation ANAMMOX in these bioreactors, respectively. Ammonia removals of 36-67% in the smart-aerated SAGBs, with measured oxygen and organic carbon limitations, further suggest partial nitritation ANAMMOX. The smart-aerated SAGBs required substantially less aeration to achieve TN removals similar to SAGBs with timer-controlled aeration. Genomic DNA testing confirmed that the dominant ANAMMOX seed bacteria, received from a treatment plant utilizing the DEMON® sidestream deammonification process, was a Candidatus Brocadia sp. (of the Planctomycetales order). The DNA from these bacteria was also present in the SAGBs at the conclusion of the study providing evidence for attached growth and limited biomass washout.

The impact of temperature on bacterial processes is well known; however temperature related data on nitrification rates in aquaculture systems are fragmented and compiled from different studies. We sought to determine ammonium and nitrite removal kinetics over a temperature range from 6 to 36 °C ...

Catalytic hydrogenation of nitrite in drinking water demands control over the selectivity towards nitrogen, minimizing the formation of ammonia. This selectivity is strongly influenced by the H/N ratio of reaction intermediates at the catalyst surface. Therefore, we fabricated a membrane reactor

Nitrate is a natural constituent of the human diet and an approved food additive. It can be partially converted to nitrogen monoxide, which induces vasodilation and thereby decreases blood pressure. This effect is associated with a reduced risk regarding cardiovascular disease, myocardial infarction, and stroke. Moreover, dietary nitrate has been associated with beneficial effects in patients with gastric ulcer, renal failure, or metabolic syndrome. Recent studies indicate that such beneficial health effects due to dietary nitrate may be achievable at intake levels resulting from the daily consumption of nitrate-rich vegetables. N-nitroso compounds are endogenously formed in humans. However, their relevance for human health has not been adequately explored up to now. Nitrate and nitrite are per se not carcinogenic, but under conditions that result in endogenous nitrosation, it cannot be excluded that ingested nitrate and nitrite may lead to an increased cancer risk and may probably be carcinogenic to humans. In this review, the known beneficial and detrimental health effects related to dietary nitrate/nitrite intake are described and the identified gaps in knowledge as well as the research needs required to perform a reliable benefit/risk assessment in terms of long-term human health consequences due to dietary nitrate/nitrite intake are presented.

This work reported new voltammetric/amperometric-based protocols using a commercial boron-doped diamond (BDD) electrode for simple and fast simultaneous detection of sulfide and nitrite from water. Square-wave voltammetry operated under the optimized working conditions of 0.01 V step potential, 0.5 V modulation amplitude and 10 Hz frequency allowed achieving the best electroanalytical parameters for the simultaneous detection of nitrite and sulfide. For practical in-field detection applications, the multiple-pulsed amperometry technique was operated under optimized conditions, i.e., -0.5 V/SCE for a duration of 0.3 s as conditioning step, +0.85 V/SCE for a duration of 3 s that assure the sulfide oxidation and +1.25 V/SCE for a duration of 0.3 s, where the nitrite oxidation occurred, which allowed the simultaneously detection of sulfide and nitrite without interference between them. Good accuracy was found for this protocol in comparison with standardized methods for each anion. Also, no interference effect was found for the cation and anion species, which are common in the water matrix.

Full Text Available that single-walled carbon nanotubes-Prussian blue hybrid (SWCNT-PB) modified electrode demonstrated greater sensitivity and catalysis towards nitrite compared to PB or a SWCNT modified electrode. The current response of the electrode was reduced...

The association between the intake of nitrate or nitrite and gastric cancer risk was investigated in a prospective cohort study started in 1986 in the Netherlands, of 120,852 men and women aged 55-69 years. At baseline, data on dietary intake, smoking habits and other covariates were collected by me

Abstract: The last two compartments of the Anaerobic Baffled Readtor ( ABR) were altered into aeration tank and sedimentation tank respectively to get an integrated anaerobic-aerobic reactor, using anaerobic granular sludge in anaerobic zone and aerobic granular sludge in aerobic zone as seed sludge. The research explored the condition to cultivate nitritation granular sludge, under the condition of continuous flow. The C/N rate was decreased from 1 to 0.4 and the ammonia nitrogen volumetric loading rate was increased from 0.89 kg x ( m3 x d)(-1) to 2.23 kg x (m3 x d)(-1) while the setting time of 1 h was controlled in the aerobic zone. After the system was operated for 45 days, the mature nitritation granular sludge in aerobic zone showed a compact structure and yellow color while the nitrite accumulation rate was about 80% in the effluent. The associated inhibition of free ammonia (FA) and free nitrous acid (FNA) dominated the nitritation. Part of granules lost stability during the initial period of operation and flocs appeared in the aerobic zone. However, the flocs were transformed into newly generated small particles in the following reactor operation, demonstrating that organic carbon was benefit to granulation and the enrichment of slow-growing nitrifying played an important role in the stability of granules.

for a few samples back in 2002. The intake, mean and intake distribution of sodium nitrite have been calculated from 1998 to 2006 with data from the Danish dietary survey conducted in 2000-02 on Danes from four to 75 years of age. The amounts used by industry have been relatively stable through the whole......The content of nitrite and nitrate in cured meat products has been monitored in Denmark seven times between 1995 and 2006. The maximum permitted added amounts of sodium nitrite in Denmark (60 mg kg(-1) for most products up to 150 mg kg(-1) for special products) have not been exceeded, except...... period with levels varying between 6 and 20 mg sodium nitrite kg(-1) with sausages, meat for open sandwiches and salami-type sausages being the greatest contributors. The mean intake of sodium nitrate was around 1 mg day(-1), which is very low compared with the total intake of 61 mg day(-1). The mean...

This article describes the reaction between nitrite and safranine O. This sensitive reaction is based on the disappearance of color of the reddish-orange azo dye, allowing the determination of nitrite at the mg mL-1 level. A factorial optimization of parameters was carried out and the method was applied for the quantification of nitrite in…

3-Oxoacyl-[ACP] reductase (E.C. 1.1.1.100, alternatively known as beta-ketoacyl-[ACP] reductase), a component of fatty acid synthetase has been purified from seeds of rape by ammonium sulphate fractionation, Procion Red H-E3B chromatography, FPLC gel filtration and high performance hydroxyapatite chromatography. The purified enzyme appears on SDS-PAGE as a number of 20-30 kDa components and has a strong tendency to exist in a dimeric form, particularly when dithiothreitol is not present to reduce disulphide bonds. Cleveland mapping and cross-reactivity with antiserum raised against avocado 3-oxoacyl-[ACP] reductase both indicate that the multiple components have similar primary structures. On gel filtration the enzyme appears to have a molecular mass of 120 kDa suggesting that the native structure is tetrameric. The enzyme has a strong preference for the acetoacetyl ester of acyl carrier protein (Km = 3 microM) over the corresponding esters of the model substrates N-acetyl cysteamine (Km = 35 mM) and CoA (Km = 261 microM). It is inactivated by dilution but this can be partly prevented by the inclusion of NADPH. Using an antiserum prepared against avocado 3-oxoacyl-[ACP] reductase, the enzyme has been visualised inside the plastids of rape embryo and leaf tissues by immunoelectron microscopy. Amino acid sequencing of two peptides prepared by digestion of the purified enzyme with trypsin showed strong similarities with 3-oxoacyl-[ACP] reductase from avocado pear and the Nod G gene product from Rhizobium meliloti.

The tolerance to ammonia and nitrites in freshwater ciliate Paramecium bursaria was measured in a conventional open system. The ciliate was exposed to different concentrations of ammonia and nitrites for 2h and 12h in order to determine the lethal concentrations. Linear regression analysis revealed that the 2h-LC50 value for ammonia was 95.94 mg/L and for nitrite 27.35 mg/L using probit scale method (with 95% confidence intervals). There was a linear correlation between the mortality probit scale and logarithmic concentration of ammonia which fit by a regression equation y=7.32x-9.51 (R2=0.98; y, mortality probit scale; x, logarithmic concentration of ammonia), by which 2 h-LC50 value for ammonia was fotnd to be 95.50 mg/L. A linear correlation between mortality probit scales and logarithmic concentration of nitrite is also followed the regression equation y=2.86x+0.89 (R2=0.95; y, mortality probit scale; x, logarithmic concentration of nitrite). The regression analysis of toxicity curves showed that the linear correlation between exposed time of ammonia-N LC50 value and ammonia-N LC50 value followed the regression equation y=2862.85e-0.0ax (R2=0.95; y, duration of exposure to LC50 value; x, LC50 value), and that between exposed time of nitrite-N LC50 value and nitrite-N LC50 value followed the regression equation y = 127.15e-0.13x (R2=0.91; y, exposed time of LC50 value; x, LC50 value). The results demonstrate that the tolerance to ammonia in P. bursaria is considerably higher than that of the larvae or juveniles of some metozoa, e.g. cultured prawns and oysters. In addition, ciliates, as bacterial predators, are likely to play a positive role in maintaining and improving water quality in aquatic environments with high-level ammonium, such as sewage treatment systems.

Full Text Available Aim: Drug-induced gingival overgrowth has a multifactorial nature and the pathogenesis is still uncertain. It has been suggested that Nitric Oxide (NO might play a role in the pathogenesis of drug-induced gingival overgrowth due to the contribution of NO to immune response and matrix degradation. NO levels in biological fluids have been used as a diagnostic biomarker in many diseases. The aim of this study is to determine whether NO levels in plasma, saliva and gingival crevicular fluid (GCF can serve as a potential biomarker for the evaluation of drug-induced gingival overgrowth risk. Material and Methods: A total of 104 patients, receiving cyclosporine A (n=35, phenytoin (n=25, nifedipine (n=26 or diltiazem (n=18 participated in the study. The amount of gingival overgrowth was evaluated with two indices and was given as percentage. Periodontal clinical parameters including plaque index (PI, gingival index (GI, gingival bleeding time index (GBTI and probing depth (PD were also assessed. Saliva, GCF and plasma samples were obtained from each participants. Nitrite and nitrate levels in saliva, GCF and plasma were analyzed by Griess reagent. Results: Salivary nitrite and nitrate levels in responders were significantly higher than those in non-responders in only phenytoin group (p˂0.05. Nitrite and nitrate levels of gingival crevicular fluid and plasma did not significantly differ between responders and non-responders in all study groups (p˃0.05. Salivary nitrite levels exhibited a significant correlation with PD, GBTI, severity of gingival overgrowth (%GO and GCF volume (p˂0.05. Additionally, a strong positive correlation was detected between saliva and plasma nitrate levels (p˂0.005. However, both nitrite and nitrate levels in GCF and plasma demonstrated no significant correlation with clinical parameters, GO severity and GCF volume (p˃0.05.Conclusion: Salivary nitrite and nitrate levels could be used as periodontal disease biomarkers in

Lignans are a widely distributed class of natural products, whose functions and distribution suggest that they are one of the earliest forms of defense to have evolved in vascular plants; some, such as podophyllotoxin and enterodiol, have important roles in cancer chemotherapy and prevention, respectively. Entry into lignan enzymology has been gained by the approximately 3000-fold purification of two isoforms of (+)-pinoresinol/(+)-lariciresinol reductase, a pivotal branchpoint enzyme in lignan biosynthesis. Both have comparable ( approximately 34.9 kDa) molecular mass and kinetic (Vmax/Km) properties and catalyze sequential, NADPH-dependent, stereospecific, hydride transfers where the incoming hydride takes up the pro-R position. The gene encoding (+)-pinoresinol/(+)-lariciresinol reductase has been cloned and the recombinant protein heterologously expressed as a functional beta-galactosidase fusion protein. Its amino acid sequence reveals a strong homology to isoflavone reductase, a key branchpoint enzyme in isoflavonoid metabolism and primarily found in the Fabaceae (angiosperms). This is of great evolutionary significance since both lignans and isoflavonoids have comparable plant defense properties, as well as similar roles as phytoestrogens. Given that lignans are widespread from primitive plants onwards, whereas the isoflavone reductase-derived isoflavonoids are mainly restricted to the Fabaceae, it is tempting to speculate that this branch of the isoflavonoid pathway arose via evolutionary divergence from that giving the lignans.

Despite the importance of plant lignans and isoflavonoids in human health protection (e.g. for both treatment and prevention of onset of various cancers) as well as in plant biology (e.g. in defense functions and in heartwood development), systematic studies on the enzymes involved in their biosynthesis have only recently begun. In this investigation, three NADPH-dependent aromatic alcohol reductases were comprehensively studied, namely pinoresinol-lariciresinol reductase (PLR), phenylcoumaran benzylic ether reductase (PCBER), and isoflavone reductase (IFR), which are involved in central steps to the various important bioactive lignans and isoflavonoids. Of particular interest was in determining how differing regio- and enantiospecificities are achieved with the different enzymes, despite each apparently going through similar enone intermediates. Initially, the three-dimensional x-ray crystal structures of both PLR_Tp1 and PCBER_Pt1 were solved and refined to 2.5 and 2.2 A resolutions, respectively. Not only do they share high gene sequence similarity, but their structures are similar, having a continuous alpha/beta NADPH-binding domain and a smaller substrate-binding domain. IFR (whose crystal structure is not yet obtained) was also compared (modeled) with PLR and PCBER and was deduced to have the same overall basic structure. The basis for the distinct enantio-specific and regio-specific reactions of PCBER, PLR, and IFR, as well as the reaction mechanism and participating residues involved (as identified by site-directed mutagenesis), are discussed.

Despite the importance of plant lignans and isoflavonoids in human health protection (e.g. for both treatment and prevention of onset of various cancers) as well as in plant biology (e.g. in defense functions and in heartwood development), systematic studies on the enzymes involved in their biosynthesis have only recently begun. In this investigation, three NADPH-dependent aromatic alcohol reductases were comprehensively studied, namely pinoresinol-lariciresinol reductase (PLR), phenylcoumaran benzylic ether reductase (PCBER), and isoflavone reductase (IFR), which are involved in central steps to the various important bioactive lignans and isoflavonoids. Of particular interest was in determining how differing regio- and enantiospecificities are achieved with the different enzymes, despite each apparently going through similar enone intermediates. Initially, the three-dimensional x-ray crystal structures of both PLR_Tp1 and PCBER_Pt1 were solved and refined to 2.5 and 2.2 A resolutions, respectively. Not only do they share high gene sequence similarity, but their structures are similar, having a continuous alpha/beta NADPH-binding domain and a smaller substrate-binding domain. IFR (whose crystal structure is not yet obtained) was also compared (modeled) with PLR and PCBER and was deduced to have the same overall basic structure. The basis for the distinct enantio-specific and regio-specific reactions of PCBER, PLR, and IFR, as well as the reaction mechanism and participating residues involved (as identified by site-directed mutagenesis), are discussed.

The heme-copper oxygen reductases are redox-driven proton pumps that generate a proton motive force in both prokaryotes and mitochondria. These enzymes have been divided into 3 evolutionarily related groups: the A-, B- and C-families. Most experimental work on proton-pumping mechanisms has been performed with members of the A-family. These enzymes require 2 proton input pathways (D- and K-channels) to transfer protons used for oxygen reduction chemistry and for proton pumping, with the D-channel transporting all pumped protons. In this work we use site-directed mutagenesis to demonstrate that the ba(3) oxygen reductase from Thermus thermophilus, a representative of the B-family, does not contain a D-channel. Rather, it utilizes only 1 proton input channel, analogous to that of the A-family K-channel, and it delivers protons to the active site for both O2 chemistry and proton pumping. Comparison of available subunit I sequences reveals that the only structural elements conserved within the oxygen reductase families that could perform these functions are active-site components, namely the covalently linked histidine-tyrosine, the Cu(B) and its ligands, and the active-site heme and its ligands. Therefore, our data suggest that all oxygen reductases perform the same chemical reactions for oxygen reduction and comprise the essential elements of the proton-pumping mechanism (e.g., the proton-loading and kinetic-gating sites). These sites, however, cannot be located within the D-channel. These results along with structural considerations point to the A-propionate region of the active-site heme and surrounding water molecules as the proton-loading site.

The production of ultrahigh purity copper (99.9999%) by electrolysis in the presence of a cementation barrier has been attempted employing a waste nitric copper etching solution as the electrolyte. The amount of copper deposited on the cathode increased almost linearly with electrolysis time and the purity of copper was observed to increase as the electrolyte concentration was increased. At some point, however, as the electrolyte concentration increased, the purity of copper decreased slightly. As the total surface area of cementation barrier increased, the purity of product increased. The electrolyte temperature should be maintained below 35 degrees C in the range of investigated electrolysis conditions to obtain the ultrahigh purity copper. Considering that several industrial waste solutions contain valuable metallic components the result of present study may support a claim that electrowinning is a very desirable process for their treatment and recovery.

Iron (Fe) and copper (Cu) homeostasis are tightly linked across biology. Understanding crosstalk between Fe and Cu nutrition could lead to strategies for improved growth on soils with low or excess metals, with implications for agriculture and phytoremediation. Here, we show that Cu and Fe nutrition interact to increase or decrease Fe and/or Cu accumulation in leaves and Fe uptake processes. Leaf Cu concentration increased under low Fe supply, while high Cu lowered leaf Fe concentration. Ferric reductase activity, an indicator of Fe demand, was inhibited at insufficient or high Cu supply. Surprisingly, plants grown without Fe were more susceptible to Cu toxicity.

Centering on the strategic goal of building "World Copper Capital", Tongling constantly extends its product lines and improves the copper industry chain. Now, the copper industry with a production value of RMB 100 billion has taken shape.As the largest copper wire rod manufacturer in Asia, Tongling Quanwei Copper Technologies Co., Ltd., upon its moving into the local market,

Full Text Available In marine systems, nitrate is the major reservoir of inorganic fixed nitrogen. The only known biological nitrate-forming reaction is nitrite oxidation, but despite its importance, our knowledge of the organisms catalyzing this key process in the marine N-cycle is very limited. The most frequently encountered marine NOB are related to Nitrospina gracilis, an aerobic chemolithoautotrophic bacterium isolated from ocean surface waters. To date, limited physiological and genomic data for this organism were available and its phylogenetic affiliation was uncertain. In this study, the draft genome sequence of Nitrospina gracilis strain 3/211 was obtained. Unexpectedly for an aerobic organism, N. gracilis lacks classical reactive oxygen defense mechanisms and uses the reductive tricarboxylic acid cycle for carbon fixation. These features indicate microaerophilic ancestry and are consistent with the presence of Nitrospina in marine oxygen minimum zones. Fixed carbon is stored intracellularly as glycogen, but genes for utilizing external organic carbon sources were not identified. N. gracilis also contains a full gene set for oxidative phosphorylation with oxygen as terminal electron acceptor and for reverse electron transport from nitrite to NADH. A novel variation of complex I may catalyze the required reverse electron flow to low-potential ferredoxin. Interestingly, comparative genomics indicated a strong evolutionary link between Nitrospina, the nitrite-oxidizing genus Nitrospira, and anaerobic ammonium oxidizers, apparently including the horizontal transfer of a periplasmically oriented nitrite oxidoreductase and other key genes for nitrite oxidation at an early evolutionary stage. Further, detailed phylogenetic analyses using concatenated marker genes provided evidence that Nitrospina forms a novel bacterial phylum, for which we propose the name Nitrospinae.

In marine systems, nitrate is the major reservoir of inorganic fixed nitrogen. The only known biological nitrate-forming reaction is nitrite oxidation, but despite its importance, our knowledge of the organisms catalyzing this key process in the marine N-cycle is very limited. The most frequently encountered marine NOB are related to Nitrospina gracilis, an aerobic chemolithoautotrophic bacterium isolated from ocean surface waters. To date, limited physiological and genomic data for this organism were available and its phylogenetic affiliation was uncertain. In this study, the draft genome sequence of N. gracilis strain 3/211 was obtained. Unexpectedly for an aerobic organism, N. gracilis lacks classical reactive oxygen defense mechanisms and uses the reductive tricarboxylic acid cycle for carbon fixation. These features indicate microaerophilic ancestry and are consistent with the presence of Nitrospina in marine oxygen minimum zones. Fixed carbon is stored intracellularly as glycogen, but genes for utilizing external organic carbon sources were not identified. N. gracilis also contains a full gene set for oxidative phosphorylation with oxygen as terminal electron acceptor and for reverse electron transport from nitrite to NADH. A novel variation of complex I may catalyze the required reverse electron flow to low-potential ferredoxin. Interestingly, comparative genomics indicated a strong evolutionary link between Nitrospina, the nitrite-oxidizing genus Nitrospira, and anaerobic ammonium oxidizers, apparently including the horizontal transfer of a periplasmically oriented nitrite oxidoreductase and other key genes for nitrite oxidation at an early evolutionary stage. Further, detailed phylogenetic analyses using concatenated marker genes provided evidence that Nitrospina forms a novel bacterial phylum, for which we propose the name Nitrospinae.

Full Text Available Partial nitrification (nitritation – anammox (anaerobic ammonia oxidation process is increasingly used to treat wastewater, characterized by a high nitrogen content and high temperature (25 - 40°C. It is connected with the optimal temperature of anammox bacteria, which is at the range between 30 and 40°C. Mainstream application of anammox for the municipal wastewater, characterized by lower temperature seems to be one of the most challenging, but profitable process. Thenceforth, the research performed in the field of the nitritation – anammox at low temperature (10 - 20°C become more and more intense. Compared with the conventional nitrification – denitrification system, nitritation – anammox reduces oxygen demand, eliminates the need for organic carbon source and produces less excess sludge. As a result, it allows to a significant cost reduction. This paper reviews the most important and recent information in the field of nitritation – anammox process at low temperature. Effective nitrogen removal from the municipal wastewater was demonstrated at 15°C in a pilot scale and at 12°C in a laboratory scale reactor. The best performance is achieved in sequencing batch reactors and moving bed reactors with biofilm or granular biomass, as well as combinations of these technologies. Molecular biology studies shows that anammox bacteria of the genus Candidatus Brocadia may have the biggest predispositions to adapt to low temperature. However, temperature about 10°C, time and method of biomass adaptation are still the main challenges for stable and common nitritation – anammox process.

Ventricina is a traditional sausage made from pork meat produced in the Abruzzi and Molise regions. The aim of this study was to detect the content of nitrates and nitrites in local cultivars of chilli pepper, and their concentration in ventricina samples spiced with the same chilli pepper. Furthermore, it was examined whether, in the samples of ventricina with nitrate addition, the spicing with chilli pepper could exceed the maximum added dose. The concentration of nitrates and nitrites in the organic chilli pepper was 531.0±94.6 mg/kg and less than 5.0 mg, respectively, in the traditional chilli pepper it was 394.0±39.6 and less than 5.0 mg, while in the commercial it was 325.0±115.0 and less than 5.0 mg. The determination of nitrites and nitrates was carried out by high performance ion chromatography. In ventricina samples produced without added sodium nitrate, nitrates and nitrites were below 5.0 mg/kg at the case-filling time (t0) and after 50 days of aging (t50). In the samples of ventricina with added sodium nitrate, nitrate concentration values were 134.0±20.9 mg/kg at t0 and 129.0±15.4 mg/kg at t50, while the nitrites were below 5.0 mg/kg at t0 and 28.8±15.8 mg/kg at t50. Although in ventricina the amount of chilli pepper is quite relevant, it did not lead to a detectable concentration of nitrates. The maximum allowed amount was never exceeded.

Full Text Available The tofu industry is one of food industry which the product of organic waste to environment pollution. One of alternative methode which used to overcome tofu industrial waste water pollution is adsorption methode using activated carbon from coffee waste. The aim of this researched is to know about the activated carbon from coffee waste quality which observe of rendemen, water content, ash content, and iodium adsorption, to know optimum contact of time and pH of coffee waste to decrease ammonia, nitrite and nitrate contents in tofu industry waste water and to know decrease percentage of ammonia, nitrite and nitrate contents in tofu industrial waste water using activated carbon from coffee waste. The activated carbon made by soaking of coffee waste in HCl 0.1 M solution for 2 days. The activated carbon coaled in muffle furnace at temperature 350°C. The activated carbon analyzed consist of rendemen, water content, ash content, and iodium adsorption. Optimum contact of time and pH of coffee waste determined in order to get optimum adsorption ammonia, nitrite and nitrate in tofu industrial waste water. Contact time variation are 1, 10, 30, 45, 60, 90, 120 minutes and pH variation are 4, 5, 6, 7, 8, 9, 10. The result showed that the activated carbon from coffee waste fulfill the criteria SNI number 06-3730-1995. The activated carbon from coffee waste could be used to decrease the ammonia, nitrite, and nitrate contents in tofu industrial waste water at the optimum contact of time of 30 minutes and pH 7. Decreasing percentage of ammonia, nitrite and nitrate contents in tofu industrial waste water are 64,69% , 52,35% and 86,40% respectively.